N1/N2-lactam acetyl-CoA carboxylase inhibitors

ABSTRACT

The invention provides a compound of Formula (I) 
                         
or a pharmaceutically acceptable salt thereof; wherein
     G is   

     
       
         
         
             
             
         
       
         
         R 1 , R 2  and R 3  are as described herein; pharmaceutical compositions thereof; and the use thereof in treating diseases, conditions or disorders modulated by the inhibition of an acetyl-CoA carboxylase enzyme(s) in an animal.

FIELD OF THE INVENTION

This invention relates to substituted pyrazolospiroketone compounds thatact as inhibitors of an acetyl-CoA carboxylase(s) and their use intreating diseases, conditions or disorders modulated by the inhibitionof acetyl-CoA carboxylase enzyme(s).

BACKGROUND OF THE INVENTION

Acetyl-CoA carboxylases (ACC) are a family of enzymes found in mostspecies and are associated with fatty acid synthesis and metabolismthrough catalyzing the production of malonyl-CoA from acetyl-CoA. Inmammals, two isoforms of the ACC enzyme have been identified. ACC1,which is expressed at high levels in lipogenic tissues, such as fat andthe liver, controls the first committed step in the biosynthesis oflong-chain fatty acids. If acetyl-CoA is not carboxylated to formmalonyl-CoA, it is metabolized through the Krebs cycle. ACC2, a minorcomponent of hepatic ACC but the predominant isoform in heart andskeletal muscle, catalyzes the production of malonyl-CoA at thecytosolic surface of mitochondria, and regulates how much fatty acid isutilized in β-oxidation by inhibiting carnitine palm itoyl transferase.Thus, by increasing fatty acid utilization and by preventing increasesin de novo fatty acid synthesis, chronic administration of an ACCinhibitor (ACC-I) may also deplete liver and adipose tissue triglyceride(TG) stores in obese subjects consuming a high or low-fat diet, leadingto selective loss of body fat.

Studies conducted by Abu-Etheiga, et al., suggest that ACC2 plays anessential role in controlling fatty acid oxidation and, as such it wouldprovide a target in therapy against obesity and obesity-relateddiseases, such as type-2 diabetes. See, Abu-Etheiga, L., et al.,“Acetyl-CoA carboxylase 2 mutant mice are protected against obesity anddiabetes induced by high-fat/high-carbohydrate diets” PNAS, 100(18)10207-10212 (2003). See also, Choi, C. S., et al., “Continuous fatoxidation in acetyl-CoA carboxylase 2 knockout mice increases totalenergy expenditure, reduces fat mass, and improves insulin sensitivity”PNAS, 104(42) 16480-16485 (2007).

It is becoming increasingly clear that hepatic lipid accumulation causeshepatic insulin resistance and contributes to the pathogenesis of type 2diabetes. Salvage, et al., demonstrated that ACC1 and ACC2 are bothinvolved in regulating fat oxidation in hepatocytes while ACC1, thedominant isoform in rat liver, is the sole regulator of fatty acidsynthesis. Furthermore, in their model, combined reduction of bothisoforms is required to significantly lower hepatic malonyl-CoA levels,increase fat oxidation in the fed state, reduce lipid accumulation, andimprove insulin action in vivo. Thus, showing that hepatic ACC1 and ACC2inhibitors may be useful in the treatment of nonalcoholic fatty liverdisease (NAFLD) and hepatic insulin resistance. See, Savage, D. B., etal., “Reversal of diet-induced hepatic steatosis and hepatic insulinresistance by antisense oligonucleotide inhibitors of acetyl-CoAcarboxylases 1 and 2” J Clin Invest doi: 10.1172/JCI27300. See also, Oh,W., et al., “Glucose and fat metabolism in adipose tissue of acetyl-CoAcarboxylase 2 knockout mice” PNAS, 102(5) 1384-1389 (2005).

Consequently, there is a need for medicaments containing ACC1 and/orACC2 inhibitors to treat obesity and obesity-related diseases (such as,NAFLD and type-2 diabetes) by inhibiting fatty acid synthesis and byincreasing fatty acid oxidation.

SUMMARY OF THE INVENTION

The present invention relates to compounds having the structure ofFormula (I)

wherein G is

R¹ is a (C₁-C₆)alkyl or (C₃-C₅) cycloalkyl; R² is phenyl, naphthyl, a 5to 12 membered heteroaryl or a 8 to 12 membered fused heterocyclicaryl;wherein each R² group is optionally substituted with one to threesubstituents independently selected from (C₁-C₃)alkyl, (C₁-C₃)alkoxyhalo and CON H₂; and R³ is hydrogen or (C₁-C₃)alkyl; or apharmaceutically acceptable salt thereof.

A preferred embodiment of the present invention are compounds of Formula(I) wherein R¹ is isopropyl or t-butyl; or a pharmaceutically acceptablesalt thereof.

Another preferred embodiment of the present invention are compounds ofFormula (I) wherein R² is benzoimidazolyl, pyrrolopyridinyl,pyrazolopyridinyl, indazolyl, quinolinyl or isoquinolinyl, said R² isoptionally mono- or di-substituted independently with one to twosubstituents independently selected from a (C₁-C₃)alkyl, (C₁-C₃)alkoxyand halo; or a pharmaceutically acceptable salt thereof. Yet anotherpreferred embodiment of the present invention are compounds of Formula(I) wherein R² is indazolyl, benzoimidazolyl, or1H-pyrrolo[3,2-b]pyridinyl, said R² isoptionally substituted with one totwo methyl, methoxy or chloro; or a pharmaceutically acceptable saltthereof.

Another preferred embodiment of the present invention is a compoundselected from1′-isopropyl-1-(2-methyl-1H-benzo[d]imidazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(2-methyl-2H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(1H-pyrrolo[2,3-b]pyridine-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;and1′-isopropyl-1-(1H-pyrrolo[3,2-b]pyridine-6-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;or a pharmaceutically acceptable salt thereof.

Another preferred embodiment of the present invention is a compoundselected from1′-isopropyl-1-(1-methyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(4,8-dimethoxyquinoline-2-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(1H-pyrrolo[3,2-b]pyridine-2-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;and1′-isopropyl-1-(1H-pyrazolo[4,3-b]pyridine-6-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;or a pharmaceutically acceptable salt thereof.

Another preferred embodiment of the present invention is a compoundselected from1-(3,7-dimethyl-1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(7-methyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-tert-butyl-1-(1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-tert-butyl-1-(7-methyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;and1′-tert-butyl-1-(3,7-dimethyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;or a pharmaceutically acceptable salt thereof.

Another preferred embodiment of the present invention is a compoundselected from1-(7-chloro-1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(4-methoxy-1H-indazole-6-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(7-ethyl-1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(3-ethyl-1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;and1′-isopropyl-1-(3-methyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;or a pharmaceutically acceptable salt thereof.

Another preferred embodiment of the present invention is a compoundselected from1-(1H-indazole-5-carbonyl)-2′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;2′-tert-butyl-1-(1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;2′-isopropyl-1-(7-methyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;1-(3,7-dimethyl-1H-indazole-5-carbonyl)-2′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;2′-tert-butyl-1-(7-methyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;2′-tert-butyl-1-(3,7-dimethyl-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;and2′-isopropyl-1-(2-methyl-1H-benzo[d]imidazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one;or a pharmaceutically acceptable salt thereof.

Another preferred embodiment of the present invention is a compoundselected from1′-isopropyl-1-(quinoline-3-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(quinoline-6-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(isoquinoline-6-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1′-isopropyl-1-(isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;and1′-isopropyl-1-(quinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is the compound of Formula(I)

or a pharmaceutically acceptable salt thereof; wherein

G is

R¹ is a (C₁-C₆)alkyl or (C₃-C₅) cycloalkyl; R² is phenyl; naphthyl; a 5to 12 membered heteroaryl or a 8 to 12 membered fused heterocyclicaryl;wherein each R² group is optionally substituted with one to threesubstituents independently selected from (C₁-C₆)alkyl,(C₃-C₇)cycloalkyl, (C₁-C₆)alkoxy, halo, cyano, CONR⁴R⁵, NR⁴R⁵, or a 3 to7 membered heterocyclyl, wherein said (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl or(C₁-C₆)alkoxy are optionally substituted with 1 to 5 fluoro; R³ ishydrogen or (C₁-C₃)alkyl; and R⁴ and R⁵ at each occurrence areindependently selected from hydrogen, (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl,(C₁-C₃)alkoxy-(C₁-C₆)alkyl or a 3 to 7 membered heterocyclyl; whereinsaid (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl or (C₁-C₃)alkoxy-(C₁-C₆)alkyl areoptionally substituted with 1 to 5 fluoro.

Yet another embodiment of the present invention is the compound ofFormula (I) wherein G is

and

R¹ is isopropyl or t-butyl; or a pharmaceutically acceptable saltthereof.

Another embodiment of the present invention is the compound of Formula(I) wherein R² is benzoimidazolyl, benzotriazolyl, pyrrolopyridinyl,pyrazolopyridinyl, indolyl, indazolyl, quinolinyl or isoquinolinyl, saidR² is optionally substituted with one to two substituents independentlyselected from (C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo or NR⁴R⁵, wherein said(C₁-C₆)alkyl or (C₁-C₆)alkoxy are optionally substituted with 1 to 5fluoro; or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is the compound of Formula(I) wherein R² is indazolyl, indolyl, benzoimidazolyl, or1H-pyrrolo[3,2-b]pyridinyl, said R² optionally substituted independentlywith one to two methyl, methoxy, NH₂, NHCH₃ or chloro; or apharmaceutically acceptable salt thereof.

Yet another embodiment of the present invention is the compound ofFormula (I) wherein R² is quinolinyl or isoquinolinyl, said R²optionally substituted independently with one to two methyl, methoxy,NH₂, NHCH₃, NHCH₂CH₃, NHCH₂CF₃ or chloro; or a pharmaceuticallyacceptable salt thereof.

A preferred embodiment of the present invention is a compound selectedfrom the group consisting of1-(3,7-dimethyl-1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(3,7-dimethyl-1H-indazole-5-carbonyl)-1′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(3,7-dimethyl-1H-indazole-5-carbonyl)-2′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(6-methoxyquinoline-3-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(1-methoxyisoquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(3-chloro-7-methyl-1H-indazole-5-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(2-methoxyquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(2-aminoquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(5-methoxyquinoline-3-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(2-amino-1H-benzo[d]imidazole-5-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(2-(methylamino)quinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(1-(methylamino)isoquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(3-chloro-1H-indole-6-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(3-chloro-1H-pyrrolo[3,2-b]pyridine-6-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(2-(methylamino)quinoline-7-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;1-(1-(2,2,2-trifluoroethylamino)quinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;and1-(1-(ethylamino)isoquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one;or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention is a pharmaceutical compositioncomprising an amount of a compound of Formula (I) as described in any ofthe embodiments; or a pharmaceutically acceptable salt thereof and apharmaceutically acceptable excipient, diluent, or carrier. Preferably,the composition comprises a therapeutically effective amount of acompound of the present invention. The composition may also contain atleast one additional pharmaceutical agent. Preferred agents includeanti-diabetic agents and/or anti-obesity agents.

In yet another aspect of the present invention is a method for treatinga disease, condition, or disorder mediated by the inhibition ofacetyl-CoA carboxylase enzyme(s) in a mammal that includes the step ofadministering to a mammal, preferably a human, in need of such treatmenta therapeutically effective amount of a compound of the presentinvention, or a pharmaceutically acceptable salt thereof or apharmaceutical composition thereof.

Diseases, disorders, or conditions mediated by inhibitors of acetyl-CoAcarboxylases include Type II diabetes and diabetes-related diseases,such as nonalcoholic fatty liver disease (NAFLD), hepatic insulinresistance, hyperglycemia, metabolic syndrome, impaired glucosetolerance, diabetic neuropathy, diabetic nephropathy, diabeticretinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, andinsulin resistance syndrome. Preferred diseases, disorders, orconditions include Type II diabetes, nonalcoholic fatty liver disease(NAFLD), hepatic insulin resistance, hyperglycemia, impaired glucosetolerance, obesity, and insulin resistance syndrome. More preferred areType II diabetes, nonalcoholic fatty liver disease (NAFLD), hepaticinsulin resistance, hyperglycemia, and obesity. Most preferred is TypeII diabetes.

A preferred embodiment is a method for treating (e.g. delaying theprogression or onset of) Type 2 diabetes and diabetes-related disordersin animals comprising the step of administering to an animal in need ofsuch treatment a therapeutically effective amount of a compound of thepresent invention or a pharmaceutically acceptable salt thereof or acomposition thereof.

Another preferred embodiment is a method for treating obesity andobesity-related disorders in animals comprising the step ofadministering to an animal in need of such treatment a therapeuticallyeffective amount of a compound of the present invention or apharmaceutically acceptable salt thereof or a composition thereof.

Yet another preferred embodiment is a method for treating nonalcoholicfatty liver disease (NAFLD) or hepatic insulin resistance in animalscomprising the step of administering to an animal in need of suchtreatment a therapeutically effective amount of a compound of thepresent invention or a pharmaceutically acceptable salt thereof or acomposition thereof.

Compounds of the present invention may be administered in combinationwith other pharmaceutical agents (in particular, anti-obesity andanti-diabetic agents described herein below). The combination therapymay be administered as (a) a single pharmaceutical composition whichcomprises a compound of the present invention, at least one additionalpharmaceutical agent described herein and a pharmaceutically acceptableexcipient, diluent, or carrier; or (b) two separate pharmaceuticalcompositions comprising (i) a first composition comprising a compound ofthe present invention and a pharmaceutically acceptable excipient,diluent, or carrier, and (ii) a second composition comprising at leastone additional pharmaceutical agent described herein and apharmaceutically acceptable excipient, diluent, or carrier. Thepharmaceutical compositions may be administered simultaneously orsequentially and in any order.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 SEQ. ID NO. 1 provides a sequence of recombinant human ACC1 (SEQ.ID NO. 1) that can be employed in the Transcreener in vitro assay.

FIG. 2 SEQ. ID NO. 2 provides a sequence of recombinant human ACC2 (SEQ.ID NO. 2) that can be employed in the Transcreener in vitro assay.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The phrase “therapeutically effective amount” means an amount of acompound of the present invention or a pharmaceutically acceptable saltthereof that: (i) treats or prevents the particular disease, condition,or disorder, (ii) attenuates, ameliorates, or eliminates one or moresymptoms of the particular disease, condition, or disorder, or (iii)prevents or delays the onset of one or more symptoms of the particulardisease, condition, or disorder described herein.

The term “animal” refers to humans (male or female), companion animals(e.g., dogs, cats and horses), food-source animals, zoo animals, marineanimals, birds and other similar animal species. “Edible animals” refersto food-source animals such as cows, pigs, sheep and poultry.

The phrase “pharmaceutically acceptable” indicates that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith.

The terms “treating”, “treat”, or “treatment” embrace both preventative,i.e., prophylactic, and palliative treatment.

The terms “modulated” or “modulating”, or “modulate(s)”, as used herein,unless otherwise indicated, refers to the inhibition of the Acetyl-CoAcarboxylases (ACC) enzyme(s) with compounds of the present invention.

The terms “mediated” or “mediating” or “mediate(s)”, as used herein,unless otherwise indicated, refers to the (i) treatment or preventionthe particular disease, condition, or disorder, (ii) attenuation,amelioration, or elimination of one or more symptoms of the particulardisease, condition, or disorder, or (iii) prevention or delay of theonset of one or more symptoms of the particular disease, condition, ordisorder described herein, by inhibiting the Acetyl-CoA carboxylases(ACC) enzyme(s). The term “compounds of the present invention” (unlessspecifically identified otherwise) refer to compounds of Formula (I) andany pharmaceutically acceptable salts of the compounds, as well as, allstereoisomers (including diastereoisomers and enantiomers), tautomers,conformational isomers, and isotopically labeled compounds. Hydrates andsolvates of the compounds of the present invention are consideredcompositions of the present invention, wherein the compound is inassociation with water or solvent, respectively.

The terms “(C₁-C₆)alkyl” and “(C₁-C₃)alkyl” are alkyl groups of thespecified number of carbons, from one to six or one to three carbons,respectively, which can be either straight chain or branched. Forexample, the term “(C₁-C₃)alkyl” has from one to three carbons andconsists of methyl, ethyl, n-propyl and isopropyl.

The term “(C₃-C₇)cycloalkyl” means a cycloalkyl group with three toseven carbon atoms and consists of cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl. The term “halo” means fluoro, chloro, bromoor iodo. The term “(C₆-C₁₀)aryl” means an aromatic carbocyclic groupconsisting of six to ten carbon atoms such as phenyl or naphthyl.

The term “5 to 12 membered heteroaryl” means a five to twelve memberedaromatic group which contains at least one heteroatom selected fromnitrogen, oxygen and sulfur. As used herein the point of attachment ofthe “5 to 12 membered heteroaryl” group is on a carbon atom of thatgroup. The “5 to 12 membered heteroaryl” group can be bicyclic.Preferred embodiments of bicyclic heteroaryls include, but are notlimited to, radicals of the following ring systems:

The term “8 to 12 membered fused heterocyclicaryl” means an 8 to 12membered ring system in which a non-aromatic heterocyclic ring is fusedto an aryl ring. As used herein the point of attachment of the “8 to 12membered fused heterocyclicaryl” group is on a carbon atom of thatgroup. The term “3 to 7 membered heterocyclyl” means a three to sevenmembered saturated ring wherein one to three of the atoms areheteroatoms selected independently from nitrogen, oxygen and sulfur.Examples of “3 to 7 membered heterocyclyl” groups include but are notlimited to groups such as aziridinyl, azetidinyl, pyrrolidinyl,piperidinyl, oxiranyl, oxetanyl, tetrahydrofuranyl,tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, piperazinyl,morpholinyl and thiomorpholinyl. The point of attachment for the “3 to 7membered heterocyclyl” can be on a carbon or nitrogen atom, asappropriate for the particular group.

In one embodiment, the compound of Formula I is a N1 lactam ACCinhibitor compound having the following structure:

wherein R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyl; R² is phenyl,naphthyl, a 5 to 12 membered heteroaryl, or a 8 to 12 membered fusedheterocyclicaryl; wherein each R² group is optionally substituted withone to three substituents independently selected from (C₁-C₃)alkyl,(C₁-C₃)alkoxy, halo and CON H₂; and R³ is hydrogen or (C₁-C₃)alkyl; or apharmaceutically acceptable salt thereof.

In one embodiment, the compound of Formula I is a N2 lactam ACCinhibitor compound having the following structure:

wherein R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyl; R² is phenyl;naphthyl; a 5 to 12 membered heteroaryl or a 8 to 12 membered fusedheterocyclicaryl; wherein each R² group is optionally substituted withone to three substituents independently selected from (C₁-C₃)alkyl,(C₁-C₃)alkoxy, halo and CON H₂; and R³ is hydrogen or (C₁-C₃)alkyl; or apharmaceutically acceptable salt thereof.

Compounds of the present invention may be synthesized by syntheticroutes that include processes analogous to those well-known in thechemical arts, particularly in light of the description containedherein. The starting materials are generally available from commercialsources such as Aldrich Chemicals (Milwaukee, Wis.) or are readilyprepared using methods well known to those skilled in the art (e.g.,prepared by methods generally described in Louis F. Fieser and MaryFieser, Reagents for Organic Synthesis, v. 1-19, Wiley, New York(1967-1999 ed.), or Beilsteins Handbuch der organischen Chemie, 4, Aufl.ed. Springer-Verlag, Berlin, including supplements (also available viathe Beilstein online database)).

For illustrative purposes, the reaction schemes depicted below providepotential routes for synthesizing the compounds of the present inventionas well as key intermediates. For a more detailed description of theindividual reaction steps, see the Examples section below. Those skilledin the art will appreciate that other synthetic routes may be used tosynthesize the inventive compounds. Although specific starting materialsand reagents are depicted in the schemes and discussed below, otherstarting materials and reagents can be easily substituted to provide avariety of derivatives and/or reaction conditions. In addition, many ofthe compounds prepared by the methods described below can be furthermodified in light of this disclosure using conventional chemistry wellknown to those skilled in the art.

In the preparation of compounds of the present invention, protection ofremote functionality (e.g., primary or secondary amine) of intermediatesmay be necessary. The need for such protection will vary depending onthe nature of the remote functionality and the conditions of thepreparation methods. Suitable amino-protecting groups (NH-Pg) includeacetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz)and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a“hydroxy-protecting group” refers to a substituent of a hydroxy groupthat blocks or protects the hydroxy functionality. Suitablehydroxyl-protecting groups (O-Pg) include for example, allyl, acetyl,silyl, benzyl, para-methoxybenzyl, trityl, and the like. The need forsuch protection is readily determined by one skilled in the art. For ageneral description of protecting groups and their use, see T. W.Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, NewYork, 1991.

The following reaction schemes, Reaction Schemes I through ReactionScheme V, provide representative procedures that are used to prepare thecompounds of Formula (I). It is to be understood that these reactionschemes are to be construed in a non-limiting manner and that reasonablevariations of the depicted methods can be used to prepare the compoundsof Formula (I).

Reaction Scheme I outlines the general procedures one could use toprovide N1 lactam ACC inhibitor compounds of the present inventionhaving Formula Ia, in which R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyland R² is phenyl, naphthyl, a 5 to 12 membered heteroaryl or a 8 to 12membered fused heterocyclicaryl; wherein each R² group is optionallysubstituted with one to three substituents independently selected from(C₁-C₃)alkyl, (C₁-C₃)alkoxy, halo and CONH₂.

According to Scheme I, the compound of formula XIa can be formed byreacting methyl 2-cyano-3-ethoxyacrylate with an appropriate alkylhydrazine (R₁NHNH₂) in the presence of a base such as potassiumcarbonate and solvent. For example, the compound of formula XIa can beformed by reacting methyl 2-cyano-3-ethoxyacrylate with an appropriatealkyl hydrazine (R₁NHNH₂,) in the presence of a base such as potassiumcarbonate (“K₂CO₃”) in refluxing ethanol to provide the desired cyclizedcompound, at a temperature of about 20° C. to about 80° C. for about 2to 24 hours.

The compound of formula Xa can be formed by converting the arylamine offormula XIa to an aryl bromide using a nitrite such as isoamylnitrite,sodium nitrite, or tert-butyl nitrite and a bromide source such ascopper(II)bromide in acetonitrile, to provide compound of formula Xa, ata temperature of about 20° C. to about 80° C. for about 2 to about 18hours.

Then, the compound of formula IXa can be prepared by treating the esterof formula Xa with a reducing agent such as diisobutylaluminium hydride(“DIBAL”) or lithium aluminum hydride (“LAH”) in an aprotic solvent suchas tetrahydrofuran (“THF”), toluene or diethyl ether at a temperature ofabout 0° C. to about 80° C. for about 1 to about 12 hours.

The compound of formula VIa can be formed by first reacting the compoundof formula IXa with a brominating agent such as phosphorus tribromide(“PBr₃”), or a mixture of carbon tetrabromide and triphenylphosphine, ata temperature of about −20° C. to about 60° C. for about 30 to about 120minutes forming the compound of formula VIIIa. The compound of formulaVIIIa is then reacted with a protected piperidine derivative compound offormula VIIa in the presence of a strong base such as lithiumbis(trimethylsilyl)amide (“LiHMDS”) or lithium diisopropylamine (“LDA”)in an aprotic solvent such as THF, toluene or diethyl ether at atemperature of about −78° C. to about 20° C. for about 1 to about 18hours. The group Pg represents an appropriate amine protecting group andis preferably N-tert-butoxycarbonyl (“BOC”) or carbobenzyloxy (“Cbz”).

Then, the compound of formula VIa is then deprotected by hydrolyzing theester group with a strong aqueous base, such as lithium hydroxide, orsodium hydroxide at a temperature of about 0° C. to about 100° C. forabout 1 to about 18 hours, forming a carboxylic acid containing compoundof formula Va.

The isocyanate compound of formula IVa can then be formed by reactingthe carboxylic acid of formula Va with diphenylphosphoryl azide (“DPPA”)in the presence of a base such as triethylamine (“Et₃N”) ordiisopropylethylamine at a temperature of about 60° C. to about 120° C.for about 1 to about 12 hours. The lactam compound of formula IIIa canthen be formed by cyclization of the isocyanate (formula IVa) using analkyl lithium, such as n-butyllithium (“n-BuLi”) or t-butyllithium(“t-BuLi”) at a temperature of about −78° C. to about 0° C. for about 5to about 120 minutes.

The lactam compound of formula (IIIa) can then be deprotected to providethe free spiropiperidine derivative of formula (IIa) using standardmethods which depend on which protecting group Pg has been employed. Forexample, when Pg represents BOC, standard strong acid deprotectionconditions, such as 4N hydrochloric acid in dioxane or trifluoroaceticacid in an appropriate solvent such as dichloromethane, can be used toremove the BOC group. When Pg represents Cbz, hydrogenation overpalladium on carbon in ethanol or treatment with a hydrogen source suchas ammonium formate or 1-methyl-1,4-cyclohexadiene in the presence ofpalladium on carbon in ethanol or ethyl acetate can be employed to carryout the deprotection.

The spiropiperidine derivative of Formula (IIa) can then be acylated byemploying standard methods to provide the compound of Formula (Ia). Forexample, the compound (la) may then be formed using a standard peptidecoupling reaction with the desired carboxylic acid (R²CO₂H). Forexample, the spiropiperidine intermediate (IIa) and carboxylic acid(R²CO₂H) may be coupled by forming an activated carboxylic acid ester,such as by contacting the carboxylic acid (R²CO₂H) with a peptidecoupling reagent, such asO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (“HATU”) or1-ethyl-3-(3-dimethyilaminopropyl)carbodiimide hydrocloride (“EDC.HCl”),in the presence or absence of an activating agent, such ashydroxybenzotriazole (“HOBt”) and in the presence of a suitable base,such as N,N-diisopropylethylamine (“DIEA”), triethylamine orN-methylmorpholine (“NMM”), in a suitable solvent such as THF and/orDMF, dimethylacetamide (“DMA”) or dichloromethane and then contactingthe activated carboxylic acid ester with the spiropiperidine derivative(IIa) to form a compound of Formula (Ia).

Reaction Scheme II outlines the general procedures one could use toprovide N2 lactam ACC inhibitor compounds of the present inventionhaving Formula Ib, in which R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyland R² is phenyl, naphthyl, a 5 to 12 membered heteroaryl or a 8 to 12membered fused heterocyclicaryl; wherein each R² group is optionallysubstituted with one to three substituents independently selected from(C₁-C₃)alkyl, (C₁-C₃)alkoxy, halo and CON H₂.

According to Scheme II, alkylation of the pyrazole compound of formulaXIb to the compound of formula X using a primary or secondary alkylhalide, such as methyl iodide, ethyl iodide, 1-bromopropane,1-iodopropane, 2-bromopropane, 2 iodopropane 1-iodobutane, 2-iodobutane,1-iodo-2-methylpropane, or 1-(bromomethyl)cyclopropane, can be carriedout in the presence of a base such as cesium carbonate (“Cs₂CO₃”) orpotassium carbonate (“K₂CO₃”) and a solvent such as dimethylformamide(“DMF”), at a temperature of about 20° C. to about 100° C. for about 1to about 12 hours.

Then, the compound of formula IXb can be prepared by treating formula Xbwith a reducing agent such as DIBAL or LAH in an aprotic solvent such asTHF, toluene, or diethyl ether, at a temperature of about −78° C. toabout 60° C. for about 1 to about 12 hours.

The compound of formula VIb can be formed by first reacting the compoundof formula IXb with a brominating agent such as PBr₃ or a mixture ofcarbon tetrabromide and triphenylphosphine, at a temperature of about−20° C. to about 60° C. for about 30 to about 120 minutes forming thecompound of formula VIIIb. The compound of formula VIIIb is then reactedwith a protected piperidine derivative compound of formula VIIb using astrong base such as lithium bis(trimethylsilyl)amide (“LiHMDS”) orlithium diisopropylamine (“LDA”) in an aprotic solvent such as THF,toluene or diethyl ether at a temperature of about −78° C. to about 20°C. for about 1 to about 18 hours. The group Pg represents an appropriateamine protecting group and is preferably BOC or Cbz.

Then, the compound formula VIb is then deprotected by hydrolyzing theester group with a strong aqueous base, such as lithium hydroxide, orsodium hydroxide at a temperature of about 0° C. to about 100° C. forabout 1 to about 18 hours, forming a carboxylic acid containing compoundof formula Vb. The isocyanate compound of formula IVb can then be formedby reacting the carboxylic acid of formula Vb with DPPA in the presenceof a base such as Et₃N or diisopropylethylamine at a temperature ofabout 60° C. to about 120° C. for about 1 to about 12 hours.

The lactam compound of formula IIIb can then be formed by cyclization ofthe isocyanate (formula IVb) using an alkyl lithium, such as n-BuLi ort-BuLi at a temperature of about −78° C. to about 0° C. for about 5 toabout 120 minutes.

The lactam compound of formula (IIIb) can then be deprotected to providethe free spiropiperidine derivative of formula (IIb) using standardmethods which depend on which protecting group Pg has been employed. Forexample, when Pg represents BOC, standard strong acid deprotectionconditions, such as 4N hydrochloric acid in dioxane or trifluoroaceticacid in an appropriate solvent such as dichloromethane, can be used toremove the BOC group. When Pg represents Cbz, hydrogenation overpalladium on carbon in ethanol or treatment with a hydrogen source suchas ammonium formate or 1-methyl-1,4-cyclohexadiene in the presence ofpalladium on carbon in ethanol or ethyl acetate can be employed to carryout the deprotection.

The spiropiperidine derivative of Formula (IIb) can then be acylated byemploying standard methods to provide the compound of Formula (Ib). Forexample, the compound (Ib) may then be formed using a standard peptidecoupling reaction with the desired carboxylic acid (R²CO₂H). Forexample, the spiropiperidine intermediate (IIb) and carboxylic acid(R²CO₂H) may be coupled by forming an activated carboxylic acid ester,such as by contacting the carboxylic acid (R²CO₂H) with a peptidecoupling reagent, such as HATU or EDC.HCl, in the presence or absence ofan activating agent, such as HOBt and in the presence of a suitablebase, such as DIEA, NMM, in a suitable solvent such as THF and/or DMF,DMA or dichloromethane and then contacting the activated carboxylic acidester with the spiropiperidine derivative (IIb) to form a compound ofFormula (Ib).

Reaction Scheme III outlines the general procedures one could use toprovide N2 lactam ACC inhibitor compounds of the present inventionhaving Formula Ic, in which R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyland R² is phenyl, naphthyl, a 5 to 12 membered heteroaryl or a 8 to 12membered fused heterocyclicaryl; wherein each R² group is optionallysubstituted with one to three substituents independently selected from(C₁-C₃)alkyl, (C₁-C₃)alkoxy, halo and CONH₂.

The lactam compound of formula IVc may be formed by cyclization of theisocyanate (formula IVa) using a strong base such as lithium2,2,6,6-tetramethylpiperidide (“LTMP”) or magnesium2,2,6,6-tetramethylpiperidide at a temperature of about −78° C. to about0° C. for about 30 minutes to about 6 hours.

The lactam compound of formula (IVc), when Pg represents BOC, may thenbe dehalogenated to provide the lactam compound of formula (IIIc) byhydrogenation in the presence of a base such as Et₃N over palladium oncarbon in ethanol or treatment with a hydrogen source such as ammoniumformate or 1-methyl-1,4-cyclohexadiene in the presence of a base such asEt₃N and palladium on carbon in ethanol or ethyl acetate at atemperature of about 20° C. to about 100° C. for about 30 minutes toabout 6 hours.

The lactam compound of formula (IIIc), when Pg represents BOC, may thenbe deprotected to provide the free spiropiperidine derivative of formula(IIc) using standard strong acid deprotection conditions, such as 4Nhydrochloric acid in dioxane or trifluoroacetic acid in an appropriatesolvent such as dichloromethane, to remove the BOC group.

The lactam compound of formula (IVc), when Pg represents Cbz, may bedehalogenated and deprotected simultaneously by hydrogenation overpalladium on carbon in ethanol or treatment with a hydrogen source suchas ammonium formate or 1-methyl-1,4-cyclohexadiene in the presence ofpalladium on carbon in ethanol or ethyl acetate.

The spiropiperidine derivative of Formula (IIc) may then be acylated byemploying standard methods to provide the compound of Formula (Ic). Forexample, the compound (Ic) may then be formed using a standard peptidecoupling reaction with the desired carboxylic acid (R²CO₂H). Forexample, the spiropiperidine intermediate (IIc) and carboxylic acid(R²CO₂H) may be coupled by forming an activated carboxylic acid ester,such as by contacting the carboxylic acid (R²CO₂H) with a peptidecoupling reagent, such as HATU or EDC.HCl, in the presence or absence ofan activating agent, such as HOBt and in the presence of a suitablebase, such as DIEA, triethylamine or NMM, in a suitable solvent such asTHF and/or DMF, DMA or dichloromethane and then contacting the activatedcarboxylic acid ester with the spiropiperidine derivative (IIc) to forma compound of Formula (Ic).

Reaction Scheme IV outlines the general procedures one could use toprovide N2 lactam ACC inhibitor compounds of the present inventionhaving Formula Id, in which R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyland R² is phenyl, naphthyl, a 5 to 12 membered heteroaryl or a 8 to 12membered fused heterocyclicaryl; wherein each R² group is optionallysubstituted with one to three substituents independently selected from(C₁-C₃)alkyl, (C₁-C₃)alkoxy, halo and CON H₂.

The lactam compound of formula IIId may be formed by palladium catalyzedcross-coupling of the bromide of formula IVc with an alkyl or alkenyltributylstannane such as methyl tri-nbutylstannane or vinyltri-nbutylstannane or allyl tri-nbutylstannane or a trialkyl boroxinesuch as trimethyl boroxine or trivinyl boroxine in the presence of apalladium catalyst such as tetrakis(triphenylphosphine)palladium(0) or aprecatalyst and ligand combination such as palladium(II)acetate and2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (“SPhos”) and in thepresence or absence of a base such as potassium carbonate in a proticsolvent such as ethanol or t-amyl alcohol or an aprotic solvent such astetrahydrofuran or dimethylformamide at a temperature of about 20° C. toabout 100° C. for about 2 hours to about 18 hours or at a temperature ofabout 100° C. to about 150° C. for about 5 minutes to about 60 minutesunder microwave heating. If a alkenyl trialkylstannane or alkenylboroxine is utilized to install the R³ group, reduction of the resultingolefin may be affected by hydrogenation over palladium on carbon inethanol or treatment with a hydrogen source such as ammonium formate or1-methyl-1,4-cyclohexadiene in the presence of palladium on carbon inethanol or ethyl acetate.

The lactam compound of formula (IIId) may then be deprotected to providethe free spiropiperidine derivative of formula (IId) using standardmethods which depend on which protecting group Pg has been employed. Forexample, when Pg represents BOC, standard strong acid deprotectionconditions. such as 4N hydrochloric acid in dioxane or trifluoroaceticacid in an appropriate solvent such as dichloromethane, can be used toremove the BOC group. When Pg represents Cbz, hydrogenation overpalladium on carbon in ethanol or treatment with a hydrogen source suchas ammonium formate or 1-methyl-1,4-cyclohexadiene in the presence ofpalladium on carbon in ethanol or ethyl acetate may be employed to carryout the deprotection.

The spiropiperidine derivative of Formula (lid) may then be acylated byemploying standard methods to provide the compound of Formula (Id). Forexample, the compound (Id) may then be formed using a standard peptidecoupling reaction with the desired carboxylic acid (R²CO₂H). Forexample, the spiropiperidine intermediate (IId) and carboxylic acid(R²CO₂H) may be coupled by forming an activated carboxylic acid ester,such as by contacting the carboxylic acid (R²CO₂H) with a peptidecoupling reagent, such as HATU or EDC.HCl, in the presence or absence ofan activating agent, such as HOBt and in the presence of a suitablebase, such as DIEA, triethylamine or NMM, in a suitable solvent such asTHF and/or DMF, DMA or dichloromethane and then contacting the activatedcarboxylic acid ester with the spiropiperidine derivative (IId) to forma compound of Formula (Id).

Reaction Scheme V outlines the general procedures one could use toprovide N2 lactam ACC inhibitor compounds of the present inventionhaving Formula Id, in which R¹ is a (C₁-C₆)alkyl or (C₃-C₅)cycloalkyland R² is phenyl, naphthyl, a 5 to 12 membered heteroaryl or a 8 to 12membered fused heterocyclicaryl; wherein each R² group is optionallysubstituted with one to three substituents independently selected from(C₁-C₃)alkyl, (C₁-C₃)alkoxy, halo and CONH₂.

According to Scheme V, the compound of formula XIe can be prepared bycondensation of a keto ester compound of formula XIIIe with anappropriate an alkyl hydrazine hydrochloride of formula XIIe such ast-butylhydrazine hydrochloride in the presence of a tertiary amine basesuch as triethylamine or N,N-diisopropylethylamine in a polar proticsolvent such as ethanol at a temperature of about 20° C. to about 100°C. for about 1 to about 12 hours.

The compound of formula Xe can be prepared by treating the compound offormula XIe with (chloromethylene)dimethylammonium chloride (VilsmeierSalt, Sigma-Aldrich, cat #280909) in a non-protic solvent such asdimethylformamide or toluene or 1,2-dichloroethane at a temperature ofabout 0° C. to about 120° C. for about 1 to 12 hours.

The compound of formula IXe can be prepared by treating the aldehyde offormula Xe with a reducing agent such as sodium borohydride in a proticsolvent such as methanol or ethanol at a temperature of about 0° C. toabout 60° C. for about 1 to about 6 hours.

The compound of formula VIe can be formed by first reacting the compoundof formula IXe with a brominating agent such as phosphorus tribromide(“PBr₃”), or a mixture of carbon tetrabromide and triphenylphosphine, ata temperature of about −20° C. to about 60° C. for about 30 to about 120minutes forming the compound of formula VIIIe. The compound of formulaVIIIe is then reacted with a protected 4-cyanopiperidine derivativecompound of formula VIIa in the presence of a strong base such aslithium bis(trimethylsilyl)amide (“LiHMDS”) or lithium diisopropylamine(“LDA”) in an aprotic solvent such as tetrahydrofuran (“THF”), tolueneor diethyl ether at a temperature of about −78° C. to about 20° C. forabout 1 to about 18 hours. The group Pg represents an appropriate amineprotecting group and is preferably N-tert-butoxycarbonyl (“BOC”) orcarbobenzyloxy (“Cbz”).

The amide compound of formula Ve can be prepared by subjecting thenitrile compound of formula VIe to hydrolysis conditions such as anaqueous hydroxide base such as lithium hydroxide or sodium hydroxide anda solvent such as methanol or ethanol or tetrahydrofuran at atemperature of about 20° C. to about 100° C. for about 1 to 12 hours.Alternatively a peroxide complex can be used such as urea-hydrogenperoxide in combination with an aqueous hydroxide base such as sodiumhydroxide in a solvent such as methanol or ethanol at a temperature ofabout 0° C. to about 60° C. for about 1 to 12 hours.

Rearrangement of the amide compound of formula Ve to the isocyanatecompound of formula IVe can be carried out by treatment with a reagentsuch as (bis(trifluoroacetoxy)iodo)benzene in the presence of aninorganic base such as sodium bicarbonate in a solvent such asacetonitrile at a temperature of about 20° C. to about 60° C. for about1 to 6 hours.

Conversion of the isocyante compound of formula IVe to the lactamcompound of formula Ille can proceed by first hydrolyzing the isocyanatein aqueous hydroxide base such as sodium hydroxide or lithium hydroxidein a solvent such as methanol or tetrahydrofuran. The resulting aminecan then be treated with an amide coupling reagent such as1-ethyl-3-(3-dimethylaminopropyl)carbodiimide or2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate with a alkyl amine base such as triethylamine orN,N-diisopropylethylamine in a solvent such as dichloromethane ordimethylformamide at a temperature of about 0° C. to about 60° C. forabout 1 to 24 hours to give the lactam compound of formula Ille.

The lactam compound of formula (IIIe) can then be deprotected to providethe free spiropiperidine derivative of formula (IIe) using standardmethods which depend on which protecting group Pg has been employed. Forexample, when Pg represents tert-butyloxycarbonyl (“BOC”) standardstrong acid deprotection conditions such as 4N hydrochloric acid indioxane or trifluoroacetic acid in an appropriate solvent such asdichloromethane can be used to remove the BOC group. When Pg representscarbobenzyloxy (“Cbz”), hydrogenation over palladium on carbon inethanol or treatment with a hydrogen source such as ammonium formate or1-methyl-1,4-cyclohexadiene in the presence of palladium on carbon inethanol or ethyl acetate can be employed to carry out the deprotection.

The spiropiperidine derivative of Formula (IIe) can then be acylated byemploying standard methods to provide the compound of Formula (Ie). Forexample, the compound (Ie) can then be formed using a standard peptidecoupling reaction with the desired carboxylic acid (R²CO₂H). Forexample, the spiropiperidine intermediate (Ile) and carboxylic acid(R²CO₂H) can be coupled by forming an activated carboxylic acid ester,such as by contacting the carboxylic acid (R²CO₂H) with a peptidecoupling reagent, such as HATU or EDC.HCl, in the presence or absence ofan activating agent, such as hydroxybenzotriazole (“HOBt”) and in thepresence of a suitable base, such as DIEA, NMM, in a suitable solventsuch as THF and/or DMF, DMA or dichloromethane and then contacting theactivated carboxylic acid ester with the spiropiperidine derivative(Ile) to form a compound of Formula (Ie).

The compounds of the present invention may be isolated and used per seor in the form of their pharmaceutically acceptable salts. In accordancewith the present invention, compounds with multiple basic nitrogen atomscan form salts with varying number of equivalents (“eq.”) of acid. Itwill be understood by practitioners that all such salts are within thescope of the present invention.

Pharmaceutically acceptable salts, as used herein in relation tocompounds of the present invention, include pharmaceutically acceptableinorganic and organic salts of the compound. These salts can be preparedin situ during the final isolation and purification of a compound, or byseparately reacting the compound thereof, with a suitable organic orinorganic acid and isolating the salt thus formed. Representative saltsinclude, but are not limited to, the hydrobromide, hydrochloride,hydroiodide, sulfate, bisulfate, nitrate, acetate, trifluoroacetate,oxalate, besylate, palmitate, pamoate, malonate, stearate, laurate,malate, borate, benzoate, lactate, phosphate, hexafluorophosphate,benzene sulfonate, tosylate, formate, citrate, maleate, fumarate,succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionateand laurylsulphonate salts, and the like. These may also include cationsbased on the alkali and alkaline earth metals, such as sodium, lithium,potassium, calcium, magnesium, and the like, as well as non-toxicammonium, quaternary ammonium, and amine cations including, but notlimited to, ammonium, tetramethylammonium, tetraethylammonium,methylammonium, dimethylammonium, trimethylammonium, triethylammonium,ethylammonium, and the like. For additional examples see, for example,Berge, et al., J. Pharm. Sci., 66, 1-19 (1977).

Compounds of the present invention may exist in more than one crystalform. Polymorphs of compounds of Formula (I) and salts thereof(including solvates and hydrates) form part of this invention and may beprepared by crystallization of a compound of the present invention underdifferent conditions. For example, using different solvents or differentsolvent mixtures for recrystallization; crystallization at differenttemperatures; various modes of cooling, ranging from very fast to veryslow cooling during crystallization. Polymorphs may also be obtained byheating or melting a compound of the present invention followed bygradual or fast cooling. The presence of polymorphs may be determined bysolid probe nuclear magnetic resonance (NMR) spectroscopy, infrared (IR)spectroscopy, differential scanning calorimetry, powder X-raydiffraction or such other techniques.

This invention also includes isotopically-labeled compounds, which areidentical to those described by Formula (I), but for the fact that oneor more atoms are replaced by an atom having an atomic mass or massnumber different from the atomic mass or mass number usually found innature. Examples of isotopes that can be incorporated into compounds ofthe invention include isotopes of hydrogen, carbon, nitrogen, oxygen,sulfur and fluorine, such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O ¹⁷O, ³⁵S, ³⁶Cl,¹²⁵I, ¹²⁹I, and ¹⁸F respectively. Certain isotopically-labeled compoundsof the present invention, for example those into which radioactiveisotopes such as ³H and ¹⁴C are incorporated, are useful in drug and/orsubstrate tissue distribution assays. Tritiated (i.e., ³H), andcarbon-14 (i.e., ¹⁴C), isotopes are particularly preferred for theirease of preparation and detectability. Further, substitution withheavier isotopes such as deuterium (i.e., ²H), can afford certaintherapeutic advantages resulting from greater metabolic stability, forexample increased in vivo half-life or reduced dosage requirements and,hence, may be preferred in some circumstances. Isotopically labeledcompounds of the present invention can generally be prepared by carryingout the procedures disclosed in the schemes and/or in the Examplesbelow, by substituting a readily available isotopically labeled reagentfor a non-isotopically labeled reagent.

The compounds of the present invention may contain stereogenic centers.These compounds may exist as mixtures of enantiomers or as pureenantiomers. Wherein a compound includes a stereogenic center, thecompounds may be resolved into the pure enantiomers by methods known tothose skilled in the art, for example by formation of diastereoisomericsalts which may be separated, for example, by crystallization; formationof stereoisomeric derivatives or complexes which may be separated, forexample, by crystallization, gas-liquid or liquid chromatography;selective reaction of one enantiomer with an enantiomer-specificreagent, for example enzymatic esterification; or gas-liquid or liquidchromatography in a chiral environment, for example on a chiral supportfor example silica with a bound chiral ligand or in the presence of achiral solvent. It will be appreciated that where the desiredstereoisomer is converted into another chemical entity by one of theseparation procedures described above, a further step is required toliberate the desired enantiomeric form. Alternatively, the specificstereoisomers may be synthesized by using an optically active startingmaterial, by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one stereoisomerinto the other by asymmetric transformation.

Compounds of the present invention may exist in different stableconformational forms which may be separable. Torsional asymmetry due torestricted rotation about an asymmetric single bond, for example becauseof steric hindrance or ring strain, may permit separation of differentconformers. The compounds of the present invention further include eachconformational isomer of compounds of Formula (I) and mixtures thereof.

Compounds of the present invention are useful for treating diseases,conditions and/or disorders modulated by the inhibition of theacetyl-CoA carboxylases enzyme(s) (in particular, ACC1 and ACC2).Another embodiment of the present invention is a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof the present invention and a pharmaceutically acceptable excipient,diluent or carrier. The compounds of the present invention (includingthe compositions and processes used therein) may also be used in themanufacture of a medicament for the therapeutic applications describedherein.

A typical formulation is prepared by mixing a compound of the presentinvention and a carrier, diluent or excipient. Suitable carriers,diluents and excipients are well known to those skilled in the art andinclude materials such as carbohydrates, waxes, water soluble and/orswellable polymers, hydrophilic or hydrophobic materials, gelatin, oils,solvents, water, and the like. The particular carrier, diluent orexcipient used will depend upon the means and purpose for which thecompound of the present invention is being applied. Solvents aregenerally selected based on solvents recognized by persons skilled inthe art as safe (GRAS) to be administered to a mammal. In general, safesolvents are non-toxic aqueous solvents such as water and othernon-toxic solvents that are soluble or miscible in water. Suitableaqueous solvents include water, ethanol, propylene glycol, polyethyleneglycols (e.g., PEG400, PEG300), etc. and mixtures thereof. Theformulations may also include one or more buffers, stabilizing agents,surfactants, wetting agents, lubricating agents, emulsifiers, suspendingagents, preservatives, antioxidants, opaquing agents, glidants,processing aids, colorants, sweeteners, perfuming agents, flavoringagents and other known additives to provide an elegant presentation ofthe drug (i.e., a compound of the present invention or pharmaceuticalcomposition thereof) or aid in the manufacturing of the pharmaceuticalproduct (i.e., for use in the preparing a medicament).

The formulations may be prepared using conventional dissolution andmixing procedures. For example, the bulk drug substance (i.e., compoundof the present invention or stabilized form of the compound (e.g.,complex with a cyclodextrin derivative or other known complexationagent)) is dissolved in a suitable solvent in the presence of one ormore of the excipients described above. The dissolution rate of poorlywater-soluble compounds may be enhanced by the use of a spray-drieddispersion, such as those described by Takeuchi, H., et al. in“Enhancement of the dissolution rate of a poorly water-soluble drug(tolbutamide) by a spray-drying solvent deposition method anddisintegrants” J. Pharm. Pharmacol., 39, 769-773 (1987); and EP0901786B1(US2002/009494), incorporated herein by reference. The compound of thepresent invention is typically formulated into pharmaceutical dosageforms to provide an easily controllable dosage of the drug and to givethe patient an elegant and easily handleable product.

The pharmaceutical compositions also include solvates and hydrates ofthe compounds of the present invention. The term “solvate” refers to amolecular complex of a compound represented by Formula (I) (includingpharmaceutically acceptable salts thereof) with one or more solventmolecules. Such solvent molecules are those commonly used in thepharmaceutical art, which are known to be innocuous to the recipient,e.g., water, ethanol, ethylene glycol, and the like, The term “hydrate”refers to the complex where the solvent molecule is water. The solvatesand/or hydrates preferably exist in crystalline form. Other solvents maybe used as intermediate solvates in the preparation of more desirablesolvates, such as methanol, methyl t-butyl ether, ethyl acetate, methylacetate, (S)-propylene glycol, (R)-propylene glycol, 1,4-butyne-diol,and the like.

The pharmaceutical composition (or formulation) for application may bepackaged in a variety of ways depending upon the method used foradministering the drug. Generally, an article for distribution includesa container having deposited therein the pharmaceutical formulation inan appropriate form. Suitable containers are well-known to those skilledin the art and include materials such as bottles (plastic and glass),sachets, ampoules, plastic bags, metal cylinders, and the like. Thecontainer may also include a tamper-proof assemblage to preventindiscreet access to the contents of the package. In addition, thecontainer has deposited thereon a label that describes the contents ofthe container. The label may also include appropriate warnings.

The present invention further provides a method of treating diseases,conditions and/or disorders modulated by the inhibition of theacetyl-CoA carboxylases enzyme(s) in an animal that includesadministering to an animal in need of such treatment a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition comprising an effective amount of a compoundof the present invention and a pharmaceutically acceptable excipient,diluent, or carrier. The method is particularly useful for treatingdiseases, conditions and/or disorders that benefit from the inhibitionof acetyl-CoA carboxylases enzyme(s).

One aspect of the present invention is the treatment of obesity, andobesity-related disorders (e.g., overweight, weight gain, or weightmaintenance).

Obesity and overweight are generally defined by body mass index (BMI),which is correlated with total body fat and estimates the relative riskof disease. BMI is calculated by weight in kilograms divided by heightin meters squared (kg/m²). Overweight is typically defined as a BMI of25-29.9 kg/m², and obesity is typically defined as a BMI of 30 kg/m².See, e.g., National Heart, Lung, and Blood Institute, ClinicalGuidelines on the Identification, Evaluation, and Treatment ofOverweight and Obesity in Adults, The Evidence Report, Washington, DC:U.S. Department of Health and Human Services, NIH publication no.98-4083 (1998).

Another aspect of the present invention is for the treatment (e.g.,delaying the progression or onset) of diabetes or diabetes-relateddisorders including Type 1 (insulin-dependent diabetes mellitus, alsoreferred to as “IDDM”) and Type 2 (noninsulin-dependent diabetesmellitus, also referred to as “NIDDM”) diabetes, impaired glucosetolerance, insulin resistance, hyperglycemia, and diabetic complications(such as atherosclerosis, coronary heart disease, stroke, peripheralvascular disease, nephropathy, hypertension, neuropathy, andretinopathy).

In yet another aspect of the present invention is the treatment ofobesity co-morbidities, such as metabolic syndrome. Metabolic syndromeincludes diseases, conditions or disorders such as dyslipidemia,hypertension, insulin resistance, diabetes (e.g., Type 2 diabetes),coronary artery disease and heart failure. For more detailed informationon Metabolic Syndrome, see, e.g., Zimmet, P. Z., et al., “The MetabolicSyndrome: Perhaps an Etiologic Mystery but Far From a Myth—Where Doesthe International Diabetes Federation Stand′?,” Diabetes &Endocrinology, 7(2), (2005); and Alberti, K. G., et al., “The MetabolicSyndrome—A New Worldwide Definition,” Lancet, 366, 1059-62 (2005).Preferably, administration of the compounds of the present inventionprovides a statistically significant (p<0.05) reduction in at least onecardiovascular disease risk factor, such as lowering of plasma leptin,C-reactive protein (CRP) and/or cholesterol, as compared to a vehiclecontrol containing no drug. The administration of compounds of thepresent invention may also provide a statistically significant (p<0.05)reduction in glucose serum levels.

In yet another aspect of the invention is the treatment of nonalcoholicfatty liver disease (NAFLD) and hepatic insulin resistance.

For a normal adult human having a body weight of about 100 kg, a dosagein the range of from about 0.001 mg to about 10 mg per kilogram bodyweight is typically sufficient, preferably from about 0.01 mg/kg toabout 5.0 mg/kg, more preferably from about 0.01 mg/kg to about 1 mg/kg.However, some variability in the general dosage range may be requireddepending upon the age and weight of the subject being treated, theintended route of administration, the particular compound beingadministered and the like. The determination of dosage ranges andoptimal dosages for a particular patient is well within the ability ofone of ordinary skill in the art having the benefit of the instantdisclosure. It is also noted that the compounds of the present inventioncan be used in sustained release, controlled release, and delayedrelease formulations, which forms are also well known to one of ordinaryskill in the art.

The compounds of this invention may also be used in conjunction withother pharmaceutical agents for the treatment of the diseases,conditions and/or disorders described herein. Therefore, methods oftreatment that include administering compounds of the present inventionin combination with other pharmaceutical agents are also provided.Suitable pharmaceutical agents that may be used in combination with thecompounds of the present invention include anti-obesity agents(including appetite suppressants), anti-diabetic agents,anti-hyperglycemic agents, lipid lowering agents, and anti-hypertensiveagents.

Suitable lipid lowering agents that can be combined with the compoundsof the present invention include, for example, those described at page30, line 20 through page 31, line 30 of WO 2011005611. The lipidlowering agents include bile acid sequestrants, HMG-CoA reductaseinhibitors, HMG-CoA synthase inhibitors, cholesterol absorptioninhibitors, acyl coenzyme A-cholesterol acyl transferase (ACAT)inhibitors, CETP inhibitors, squalene synthetase inhibitors, PPAR αagonists, FXR receptor modulators, LXR receptor modulators, lipoproteinsynthesis inhibitors, rennin angiotensisn system inhibitors, PPAR δpartial agonists, bile acid reabsorption inhibitors, PPAR α agonists,triglyceride synthesis inhibitors, microsomal triglyceride transportinhibitors, transcription modulators, squalene epoxidase inhibitors, lowdensity lipoprotein receptor inducers, platelet aggregation inhibitors,5-LO or FLAP inhibitors, niacin bound chromium and other agents thataffect lipid composition.

Suitable anti-hypertensive agents that can be combined with thecompounds of the present invention include, for example, those describedat page 31, line 31 through page 32, line 18 of WO 2011005611. Theanti-hypertensive agents include diuretics, beta-adrenergic blockers,calcium channel blockers, angiotensin converting enzyme (ACE)inhibitors, neutral endopeptidase inhibitors, endothelin antagonists,vasodilators, angiotensin II receptor antagonists, α/β adrenergicblockers, alpha 1 blockers, alpha 2 agonists, aldosterone inhibitors,mineraocorticoid receptor inhibitors, renin inhibitors andangiopoietin-2-binding agents.

Suitable anti-diabetic agents include an acetyl-CoA carboxylase-(ACC)inhibitor such as those described in WO2009144554, WO2003072197,WO2009144555 and WO2008065508, a diacylglycerol O-acyltransferase 1(DGAT-1) inhibitor, such as those described in WO09016462 orWO2010086820, AZD7687 or LCQ908, diacylglycerol O-acyltransferase 2(DGAT-2) inhibitor, monoacylglycerol O-acyltransferase inhibitors, aphosphodiesterase (PDE)-10 inhibitor, an AMPK activator, a sulfonylurea(e.g., acetohexamide, chlorpropamide, diabinese, glibenclamide,glipizide, glyburide, glimepiride, gliclazide, glipentide, gliquidone,glisolamide, tolazamide, and tolbutamide), a meglitinide, an α-amylaseinhibitor (e.g., tendamistat, trestatin and AL-3688), an α-glucosidehydrolase inhibitor (e.g., acarbose), an α-glucosidase inhibitor (e.g.,adiposine, camiglibose, emiglitate, miglitol, voglibose, pradimicin-Q,and salbostatin), a PPARγ agonist (e.g., balaglitazone, ciglitazone,darglitazone, englitazone, isaglitazone, pioglitazone, rosiglitazone andtroglitazone), a PPAR α/γ agonist (e.g., CLX-0940, GW-1536, GW-1929,GW-2433, KRP-297, L-796449, LR-90, MK-0767 and SB-219994), a biguanide(e.g., metformin), a glucagon-like peptide 1 (GLP-1) modulator such asan agonist (e.g., exendin-3 and exendin-4), liraglutide, albiglutide,exenatide (Byetta®), albiglutide, taspoglutide, lixisenatide,dulaglutide, semaglutide, NN-9924,TTP-054, a protein tyrosinephosphatase-1B (PTP-1B) inhibitor (e.g., trodusquemine, hyrtiosalextract, and compounds disclosed by Zhang, S., et al., Drug DiscoveryToday, 12(9/10), 373-381 (2007)), SIRT-1 inhibitor (e.g., resveratrol,GSK2245840 or GSK184072), a dipeptidyl peptidease IV (DPP-IV) inhibitor(e.g., those in WO2005116014, sitagliptin, vildagliptin, alogliptin,dutogliptin, linagliptin and saxagliptin), an insulin secreatagogue, afatty acid oxidation inhibitor, an A2 antagonist, a c-jun amino-terminalkinase (JNK) inhibitor, glucokinase activators (GKa) such as thosedescribed in WO2010103437, WO2010103438, WO2010013161, WO2007122482,TTP-399, TTP-355, TTP-547, AZD1656, ARRY403, MK-0599, TAK-329, AZD5658or GKM-001, insulin, an insulin mimetic, a glycogen phosphorylaseinhibitor (e.g. GSK1362885), a VPAC2 receptor agonist, SGLT2 inhibitors,such as those described in E. C. Chao et al. Nature Reviews DrugDiscovery 9, 551-559 (July 2010) including dapagliflozin, canagliflozin,BI-10733, tofogliflozin (CSG452), ASP-1941, THR1474, TS-071, ISIS388626and LX4211 as well as those in WO2010023594, a glucagon receptormodulator such as those described in Demong, D. E. et al. Annual Reportsin Medicinal Chemistry 2008, 43, 119-137, GPR119 modulators,particularly agonists, such as those described in WO2010140092,WO2010128425, WO2010128414, WO2010106457, Jones, R. M. et al. inMedicinal Chemistry 2009, 44, 149-170 (e.g. MBX-2982, GSK1292263, APD597and PSN821), FGF21 derivatives or analogs such as those described inKharitonenkov, A. et al. et al., Current Opinion in InvestigationalDrugs 2009, 10(4)359-364, TGR5 (also termed GPBAR1) receptor modulators,particularly agonists, such as those described in Zhong, M., CurrentTopics in Medicinal Chemistry, 2010, 10(4), 386-396 and INT777, GPR40agonists, such as those described in Medina, J. C., Annual Reports inMedicinal Chemistry, 2008, 43, 75-85, including but not limited toTAK-875, GPR120 modulators, particularly agonists, high affinitynicotinic acid receptor (HM74A) activators, and SGLT1 inhibitors, suchas GSK1614235. A further representative listing of anti-diabetic agentsthat can be combined with the compounds of the present invention can befound, for example, at page 28, line 35 through page 30, line 19 ofWO2011005611. Preferred anti-diabetic agents are metformin and DPP-IVinhibitors (e.g., sitagliptin, vildagliptin, alogliptin, dutogliptin,linagliptin and saxagliptin). Other antidiabetic agents could includeinhibitors or modulators of carnitine palmitoyl transferase enzymes,inhibitors of fructose 1,6-diphosphatase, inhibitors of aldosereductase, mineralocorticoid receptor inhibitors, inhibitors of TORC2,inhibitors of CCR2 and/or CCR5, inhibitors of PKC isoforms (e.g. PKCα,PKCβ, PKC), inhibitors of fatty acid synthetase, inhibitors of serinepalmitoyl transferase, modulators of GPR81, GPR39, GPR43, GPR41, GPR105,Kv1.3, retinol binding protein 4, glucocorticoid receptor, somatostainreceptors (e.g. SSTR1, SSTR2, SSTR3 and SSTR5), inhibitors or modulatorsof PDHK2 or PDHK4, inhibitors of MAP4K4, modulators of IL1 familyincluding IL1 beta, modulators of RXRalpha. In addition suitableanti-diabetic agents include mechanisms listed by Carpino, P. A.,Goodwin, B. Expert Opin. Ther. Pat, 2010, 20(12), 1627-51.

Suitable anti-obesity agents (some of which may also act asanti-diabetic agents as well) include 11β-hydroxy steroiddehydrogenase-1 (11β-HSD type 1) inhibitors, stearoyl-CoA desaturase-1(SCD-1) inhibitor, MCR-4 agonists, cholecystokinin-A (CCK-A) agonists,monoamine reuptake inhibitors (such as sibutramine), sympathomimeticagents, β₃ adrenergic agonists, dopamine agonists (such asbromocriptine), melanocyte-stimulating hormone analogs, 5HT2c agonists,melanin concentrating hormone antagonists, leptin (the OB protein),leptin analogs, leptin agonists, galanin antagonists, lipase inhibitors(such as tetrahydrolipstatin, i.e. orlistat), anorectic agents (such asa bombesin agonist), neuropeptide-Y antagonists (e.g., NPY Y5antagonists such as velneperit), PYY₃₋₃₆ (including analogs thereof),BRS3 modulator, mixed antagonists of opiod receptor subtypes,thyromimetic agents, dehydroepiandrosterone or an analog thereof,glucocorticoid agonists or antagonists, orexin antagonists,glucagon-like peptide-1 agonists, ciliary neurotrophic factors (such asAxokine™ available from Regeneron Pharmaceuticals, Inc., Tarrytown, N.Y.and Procter & Gamble Company, Cincinnati, Ohio), human agouti-relatedprotein (AGRP) inhibitors, histamine 3 antagonists or inverse agonists,neuromedin U agonists, MTP/ApoB inhibitors (e.g., gut-selective MTPinhibitors, such as dirlotapide, JTT130, Usistapide, SLx4090), opioidantagonist, mu opioid receptor modulators, including but not limited toGSK1521498, MetAp2 inhibitors, including but not limited to ZGN-433,agents with mixed modulatory activity at 2 or more of glucagon, GIP andGLP1 receptors, such as MAR-701 or ZP2929, norepinephrine transporterinhibitors, cannabinoid-1-receptor antagonist/inverse agonists, ghrelinagonists/antagonists, oxyntomodulin and analogs, monoamine uptakeinhibitors, such as but not limited to tesofensine, an orexinantagonist, combination agents (such as bupropion plus zonisamide,pramlintide plus metreleptin, bupropion plus naltrexone, phentermineplus topiramate), and the like.

Preferred anti-obesity agents for use in the combination aspects of thepresent invention include gut-selective MTP inhibitors (e.g.,dirlotapide, mitratapide and implitapide, R56918 (CAS No. 403987) andCAS No. 913541-47-6), CCKa agonists (e.g.,N-benzyl-2-[4-(1H-indol-3-ylmethyl)-5-oxo-1-phenyl-4,5-dihydro-2,3,6,10b-tetraaza-benzo[e]azulen-6-yl]-N-isopropyl-acetamidedescribed in PCT Publication No. WO 2005/116034 or US Publication No.2005-0267100 A1), 5HT2c agonists (e.g., lorcaserin), MCR4 agonist (e.g.,compounds described in U.S. Pat. No. 6,818,658), lipase inhibitor (e.g.,Cetilistat), PYY₃₋₃₆ (as used herein “PYY₃₋₃₆” includes analogs, such aspeglated PYY₃₋₃₆ e.g., those described in US Publication 2006/0178501),opioid antagonists (e.g., naltrexone), oleoyl-estrone (CAS No.180003-17-2), obinepitide (TM30338), pramlintide (Symlin®), tesofensine(NS2330), leptin, bromocriptine, orlistat, AOD-9604 (CAS No.221231-10-3) and sibutramine. Preferably, compounds of the presentinvention and combination therapies are administered in conjunction withexercise and a sensible diet.

All of the recited U.S. patents and publications (including alltechnical bulletins referenced in the Examples) are incorporated hereinby reference in their entireties.

The Examples set forth herein below are for illustrative purposes only.The compositions, methods, and various parameters reflected herein areintended only to exemplify various aspects and embodiments of theinvention, and are not intended to limit the scope of the claimedinvention in any way.

The preparations described below were used in the synthesis of compoundsexemplified in the following examples.

The following commercially available starting materials were used toprepare compounds described in the Examples below: methyl3-iodo-1H-indazole-5-carboxylate (Anichem LLC, North Brunswick, N.J.),(1R,5S)-8-(tert-butoxycarbonyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid (AstaTech, Inc., Bristol, Pa.), 6-bromoisoquinolin-3-amine (ArkPharm, Inc., Libertyville, Ill.), 3-hydroxy-1H-indazole-5-carboxylicacid (Aces Pharma, Inc., Branford, Conn.), ethyl quinoline-7-carboxylate(ASW MedChem, Inc., New Brunswick, N.J.), 7-bromoisoquinolin-1(2H)-one(Alfa Aesar, Ward Hill, Mass.),3-oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (ASW MedChem, Inc., NewBrunswick, N.J.), 5-bromo-3-(trifluoromethyl)-1H-indazole (J&W Pharm LabLLC., Levittown, Pa.), 6-bromoisoquinolin-1(2H)-one (Anichem LLC, NorthBrunswick, N.J.), methyl 1H-pyrrolo[3,2-b]pyridine-6-carboxylate (ACSScientific Inc., Metuchen, N.J.), 4-bromo-2-fluoro-N-methylbenzamide(Oakwood Products, Inc., West Columbia, S.C.),7-bromo-3-chloroisoquinoline (Allichem LLC, Baltimore, Md.),7-bromoisoquinolin-3-amine (Allichem LLC, Baltimore, Md.),6-bromoisoquinolin-3-ol (Ark Pharm, Inc., Libertyville, Ill.),1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid (ASDI Inc., Newark, Del.),1-chloroisoquinoline-7-carboxylic acid (American Custom Chemicals Corp.,San Diego, Calif.), 3,7-dimethyl-1H-indazole-5-carboxylic acid (AnnkerOrganics Co. Ltd., Wuhan, China), 7-methyl-1H-indazole-5-carboxylic acid(J & W PharmLab LLC, Levittown, Pa.), 2-methyl-2H-indazole-5-carboxylicacid (Bepharm Ltd., Shanghai, China),1H-pyrrolo[3,2-b]pyridine-6-carboxylic acid (Sinova Inc., Bethesda,Md.), 7-chloro-1H-indazole-5-carboxylic acid (Annker Organics Co. Ltd.,Wuhan, China), 4-methoxy-1H-indazole-6-carboxylic acid (ASW MedChemInc., New Brunswick, N.J.), 1-methyl-1H-indazole-5-carboxylic acid (J &W PharmLab LLC, Levittown, Pa.), 7-ethyl-1H-indazole-5-carboxylic acid(Annker Organics Co. Ltd., Wuhan, China),3-ethyl-1H-indazole-5-carboxylic acid (Allichem LLC, Baltimore, Md.),3-methyl-1H-indazole-5-carboxylic acid (Ark Pharm Inc., Libertyville,Ill.), 1H-pyrrolo[3,2-b]pyridine-2-carboxylic acid (Aces Pharma Inc.,Branford, Conn.), quinoline-3-carboxylic acid (Beta Pharma Inc.,Branford, Conn.), quinoline-7-carboxylic acid (Ark Pharm Inc.,Libertyville, Ill.), isoquinoline-6-carboxylic acid (Ark Pharm Inc.,Libertyville, Ill.), isoquinoline-7-carboxylic acid (Indofine ChemicalCompany Inc., Hillsborough, N.J.), 6-methoxyquinoline-3-carboxylic acid(Princeton Biomolecular Research Inc., Monmouth Junction, N.J.),4-methoxy-7-methyl-1H-indole-2-carboxylic acid (Aurora Fine ChemicalsLLC, San Diego, Calif.), 2-aminoquinoline-6-carboxylic acid (PrincetonBiomolecular Research Inc., Monmouth Junction, N.J.),8-methoxyquinoline-3-carboxylic acid (BioBlocks Inc., San Diego,Calif.), 2-aminoquinoline-7-carboxylic acid (Princeton BiomolecularResearch Inc., Monmouth Junction, N.J.),2-methyl-1H-benzo[d]imidazole-5-carboxylic acid (Acros Organics, Geel,Belgium), 1H-indazole-5-carboxylic acid (Sigma Aldrich, St. Louis, Mo.),quinoline-6-carboxylic acid (Acros Organics, Geel, Belgium),6-methoxy-2-naphthoic acid (Sigma Aldrich, St. Louis, Mo.),1H-indazole-6-carboxylic acid (Sigma Aldrich, St. Louis, Mo.),1H-benzo[d][1,2,3]triazole-5-carboxylic acid (Sigma Aldrich, St. Louis,Mo.), 3,4-diamino-5-chlorobenzoic acid (Princeton BioMolecular Research,Inc., Monmouth Junction, N.J.), 7-bromo-1-chloroisoquinoline (AlfaAesar, Ward Hill, Mass.) 7-bromoquinoline (Anichem LLC, North Brunswick,N.J.).

The following carboxylic acids (which were used to prepare compoundsdescribed in the Examples below) were prepared by previously publishedmeans: 3,7-dimethyl-1H-indazole-5-carboxylic acid (PCT Publication No.WO2009144554), 7-methyl-1H-indazole-5-carboxylic acid (PCT PublicationNo. WO2009144554), 7-methoxy-2-naphthoic acid (PCT Publication No.WO2003018586), 5-methoxy-2-naphthoic acid (PCT Publication No.WO2003072578), 4,8-dim ethoxyquinoline-2-carboxylic acid (PCTPublication No. WO2007011809),3-chloro-7-methyl-1H-indazole-5-carboxylic acid (PCT Publication No.WO2009144554), 3-chloro-1H-indazole-5-carboxylic acid (PCT PublicationNo. WO2009144554), 8-methoxy-2-naphthoic acid (PCT Publication No.WO2003072578), 3-chloro-1H-indole-5-carboxylic acid (PCT Publication No.WO2008065508), 3-chloro-1H-indole-6-carboxylic acid (PCT Publication No.WO2008065508), 7-methoxy-3-methyl-1H-indazole-5-carboxylic acid(WO2009144554), 4,8-dimethoxyquinoline-2-carboxylic acid (PCTPublication No. WO2007011809).

EXAMPLES

The compounds and intermediates described below were named using thenaming convention provided with Chemdraw Ultra, Version 11.0.1(CambridgeSoft Corp., Cambridge Mass.). The naming convention providedwith Chemdraw Ultra, Version 11.0.1 are well known by those skilled inthe art and it is believed that the naming convention provided withChemdraw Ultra, Version 11.0.1 generally comports with the IUPAC(International Union for Pure and Applied Chemistry) recommendations onNomenclature of Organic Chemistry and the CAS Index rules. Unless notedotherwise, all reactants were obtained commercially. All of thereferences cited herein below are incorporated by reference.

Flash chromatography was performed according to the method described byStill et al., J. Org. Chem., 1978, 43, 2923.

All Biotage® purifications, discussed herein, were performed usingBiotage® SNAP columns containing KP-SIL silica (40-63 μM, 60 Angstroms)(Biotage AB; Uppsala, Sweden).

All Combiflash® purifications, discussed herein, were performed using aCombiFlash® Companion system (Teledyne Isco; Lincoln, Nebr.) utilizingpacked RediSep® silica columns

Mass Spectra were recorded on a Waters (Waters Corp.; Milford, Mass.)Micromass Platform II spectrometer. Unless otherwise specified, massspectra were recorded on a Waters (Milford, Mass.) Micromass Platform IIspectrometer.

Proton NMR chemical shifts are given in parts per million downfield fromtetramethylsilane and were recorded on a Varian Unity 300, 400 or 500MHz (megaHertz) spectrometer (Varian Inc.; Palo Alto, Calif.). NMRchemical shifts are given in parts per million downfield fromtetramethylsilane (for proton) or fluorotrichloromethane (for fluorine).

HPLC retention times were measured using the following methods: MethodA: column: Waters Atlantis dC18 4.6×50 mm, 5 μm; mobile phase A: 0.05%TFA in water (v/v); mobile phase B: 0.05% TFA in acetonitrile (v/v);gradient: 95% N5% B linear to 5% A/95% B in 4.0 minutes, hold at 5% N95%B to 5.0 minutes; flow rate: 2.0 mL/minute. Method B: column: WatersXBridge C18 4.6×50 mm, 5 μm; mobile phase A: 0.03% NH₄OH in water (v/v);mobile phase B: 0.03% NH₄OH in acetonitrile (v/v); gradient: 95% N5% Blinear to 5% A/95% B in 4.0 minutes, hold at 5% A/95% B to 5.0 minutes;flow rate: 2.0 mL/minute.

The preparations described below were used in the synthesis of compoundsexemplified in the following examples.

Preparation of Intermediates and Starting Materials

Carboxylic acid intermediates were purchased commercially, prepared asdescribed below, prepared as described in PCT Publication No. WO2009/144554, prepared using preparations well-known to those skilled inthe art, or prepared in a manner analogous to routes described above forother carboxylic acid intermediates.

Intermediate 1:1′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-onehydrochloride salt

Step1. ethyl 5-bromo-1-tert-butyl-1H-pyrazole-4-carboxylate

To a solution of ethyl 5-amino-1-tert-butyl-1H-pyrazole-4-carboxylate(674 mg, 3.19 mmol, Li et al. J. Heterocycl. Chem., 2007, 44, 749) inacetonitrile (20 mL) were added copper(II)bromide (720 mg, 3.19 mmol)and isoamylnitrite (0.56 mL, 4.15 mmol). The golden suspension washeated at 45° C. for 2 hours and then cooled to room temperature,diluted with ethyl acetate (100 mL) and washed with saturated aqueoussodium bicarbonate (50 mL), water (50 mL) and brine (50 mL). The organicphase was dried over sodium sulfate, filtered and concentrated underreduced pressure. The resultant residue was purified by flashchromatography (5-40% ethyl acetate/heptanes, 10 g silica gel) to yield685 mg of ethyl 5-bromo-1-tert-butyl-1H-pyrazole-4-carboxylate as aclear oil. +APCI (M+H) 275.0; ¹H NMR (400 MHz, CDCl₃, δ): 7.87 (s, 1H),4.32 (q, J=7.0 Hz, 2H), 1.77 (s, 9H), 1.36 (t, J=7.1 Hz, 3H).

Step 2: (5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methanol

A solution of ethyl 5-bromo-1-tert-butyl-1H-pyrazole-4-carboxylate (685mg, 2.49 mmol) in THF (20 mL) was cooled to −78° C. and treated withdiisobutylaluminum hydride (7.47 mL, 7.47 mmol, 1 M THF), dropwise. Themixture was stirred at −78° C. for 30 minutes and then warmed to roomtemperature for 18 hours. The mixture was quenched with ethyl acetate 10mL) and stirred 15 minutes. The mixture was then treated with saturatedaqueous Rochelle's salt (25 mL) and stirred 1 hour at room temperature.The mixture was diluted with ethyl acetate (100 mL) and washed withwater (100 mL). The organic layer was dried over sodium sulfate,filtered and concentrated. The residue was purified by flashchromatography (10-80% ethyl acetate/heptane gradient, 25 g silica gel)to yield 460 mg of (5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methanol as aclear oil. +APCI (M+H) 233.1, (M+2+H) 235.1; ¹H NMR (400 MHz, CDCl₃, δ):7.51 (s, 1H), 4.53 (d, 2H), 1.74 (s, 9H), 1.55 (t, J=5.8 Hz, 1H).

Step 3: 5-bromo-4-(bromomethyl)-1-tert-butyl-1H-pyrazole

A solution of (5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methanol (460 mg,1.97 mmol) in dichloromethane (25 mL) was cooled to 0° C. and thentreated with phosphorus(III)bromide (0.37 mL, 3.46 mmol), dropwise, over5 minutes. The mixture was stirred 30 minutes at 0° C. and then 1 hourat room temp. The mixture was quenched slowly with water (50 mL),stirred 30 minutes, and then extracted with ethyl acetate (2×50 mL). Theorganic phase was washed with saturated aqueous sodium bicarbonate (50mL), dried over sodium sulfate, filtered and concentrated to yield 362mg of 5-bromo-4-(bromomethyl)-1-tert-butyl-1H-pyrazole as a clear oil.¹H NMR (400 MHz, CDCl₃, δ): 7.54 (s, 1H), 4.39 (s, 2H), 1.74 (s, 9H).

Step 4: 1-tert-butyl 4-ethyl4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate

A solution of 1-tert-butyl 4-ethyl piperidine-1,4-dicarboxylate (0.37mL, 1.47 mmol) in THF (15 mL) was cooled to −78° C. and then treatedwith lithium bis(trimethylsilyl) amide (1.48 mL, 1.48 mmol, 1 Mtoluene), dropwise. The reaction was stirred 15 minutes at −78° C.,warmed to 0° C. for 30 minutes and then cooled back to −78° C. Asolution of 5-bromo-4-(bromomethyl)-1-tert-butyl-1H-pyrazole (335 mg,1.13 mmol) in THF (10 mL) was added, the mixture was stirred 1 hour at−78° C., and then allowed to stir 18 hours at room temperature. Thereaction was quenched with saturated aqueous ammonium chloride (20 mL),stirred 30 minutes at room temperature, diluted with water (50 mL) andextracted with ethyl acetate (2×50 mL). The organics were combined,dried over sodium sulfate, filtered and concentrated. The resultantresidue was purified by flash chromatography (5-40% ethylacetate/heptane, 25 g silica gel) to yield 256 mg of 1-tert-butyl4-ethyl4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylateas a clear oil. +ESI (M+H) 474.2, (M+2+H) 476.2; ¹H NMR (400 MHz, CDCl₃,δ): 7.20 (s, 1H), 4.16 (q, J=7.2 Hz, 2H), 3.93 (br. s., 2H), 2.84 (m,2H), 2.66 (s, 2H), 2.10 (d, J=12.5 Hz, 2H), 1.72 (s, 9H), 1.45 (m, 11H),1.25 (t, J=7.1 Hz, 3H).

Step 5:4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylicacid

To a solution of 1-tert-butyl 4-ethyl4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate(256 mg, 0.54 mmol) in methanol (15 mL) was added aqueous 2.5 M NaOH (5mL), and the resultant mixture was heated at reflux for 18 hours. Themixture was cooled to room temperature and methanol was removed underreduced pressure. The resultant slurry was taken up in 25 mL water,acidified with aqueous 1 N HCl, and then extracted with ethyl acetate(2×50 mL). The combined organics were dried over sodium sulfate,filtered and concentrated to yield 241 mg of4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylicacid as a colorless solid. +APCI (M+H) 444.2, (M+2+H) 446.2; ¹H NMR (400MHz, CDCl₃, δ): 7.35 (s, 1H), 3.95 (br. s., 2H), 2.92 (br. s., 2H), 2.71(s, 2H), 2.08 (d, J=12.9 Hz, 2H), 1.73 (s, 9H), 1.50 (m, 11H).

Step 6: tert-butyl4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate

To a solution of4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylicacid (241 mg, 0.54 mmol) in toluene (10 mL) was added triethylamine (91μL, 0.65 mmol) and diphenylphosphoryl azide (0.14 mL, 0.65 mmol). Themixture was heated at 120° C. for 3 hours, the reaction was cooled andthe volatiles were removed under reduced pressure. The resultant oil waspurified by flash chromatography (25 g silica, 7-60% ethylacetate/heptane gradient) to yield 225 mg of tert-butyl4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylateas a clear oil. +APCI (M+H) 385.1; ¹H NMR (400 MHz, CDCl₃, δ): 7.40 (s,1H), 4.03 (br. s., 2H), 2.97 (br. t, J=12.3, 12.3 Hz, 2H), 2.70 (s, 2H),1.74 (s, 9H), 1.67 (m, 2H), 1.62 (m, 2H), 1.46 (s, 9H).

Step 7: tert-butyl1′-tert-butyl-7′-oxo-1′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate

A solution of tert-butyl4-((5-bromo-1-tert-butyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate(225 mg, 0.51 mmol) in THF (10 mL) was cooled to −78° C. and t-butyllithium (0.6 mL, 1.7 M in pentane) was added, dropwise, over 2 minutes.The mixture was stirred 30 minutes at −78° C., warmed to 0° C., and thenquenched with saturated aqueous NH₄Cl (20 mL). The mixture was stirred30 minutes at room temperature, diluted with water (25 mL), and thenextracted with ethyl acetate (2×50 mL). The combined organics were driedover sodium sulfate, filtered and concentrated. The residue was purifiedby flash chromatography (12-100% ethyl acetate/heptane, 10 g silica gel)to yield 137 mg of tert-butyl1′-tert-butyl-7′-oxo-1′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylateas a colorless solid. +ESI (M-tBu) 307.2; ¹H NMR (400 MHz, DMSO-d₆, δ):7.74 (s, 1H), 7.30 (s, 1H), 3.51 (m, 2H), 3.20 (m, 2H), 2.79 (s, 2H),1.64 (s, 9H), 1.56 (t, J=5.8 Hz, 4H), 1.38 (s, 9H).

Step 8:1′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-onehydrochloride salt

To a solution of tert-butyl1′-tert-butyl-7′-oxo-1′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(137 mg, 0.39 mmol) in ethyl acetate (4 mL) was added 4 N HCl in dioxane(2 mL). After stirring 1 hour at room temperature, the volatiles wereremoved under reduced pressure and the resultant colorless solid wastriturated from heptane (10 mL) to yield 112 mg of the title compound asa colorless solid. +APCI (M+H) 263.3; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.84(m, 2H), 8.00 (s, 1H), 7.29 (s, 1H), 3.13 (d, J=6.1 Hz, 2H), 3.03 (br.s., 2H), 2.78 (s, 2H), 1.76 (m, 4H), 1.60 (s, 9H).

Intermediate 2:1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-oneshown below, was prepared as follows

Step 1: 5-amino-1-isopropyl-1H-pyrazole-4-carboxylate

A mixture of ethyl 2-cyano-3-ethoxyacrylate (84.4 g, 0.50 mol),isopropyl hydrazine hydrochloride (55.2 g, 0.50 mol) and potassiumcarbonate (68.8 g, 0.50 mol) in 90% ethanol/methanol (1.5 L) was heatedunder reflux for 16 hours. The solvent was then removed in vacuo andwater and ethyl acetate were added. The mixture was separated and theorganic layer was dried over magnesium sulfate, filtered and the solventwas removed in vacuo to yield ethyl5-amino-1-isopropyl-1H-pyrazole-4-carboxylate (92.4 g, 94%). +ESI (M+H)198.1; ¹H NMR (400 MHz, CDCl₃, δ): 7.63 (s, 1H), 4.97 (br.s., 2H), 4.28(q, 2H), 4.18 (m, 1H), 1.45 (d, 6H), 1.31 (t, 3H).

Step 2: 5-amino-1-isopropyl-1H-pyrazole-4-carboxylate

To a mixture of ethyl 5-amino-1-isopropyl-1H-pyrazole-4-carboxylate(107.5 g, 0.55 mol) in acetonitrile (1 L) was added copper (II) bromide(182.6 g, 0.82 mol) at room temperature, under argon. The mixture washeated to 50° C. and isoamyl nitrite (109.8 mL, 0.82 mol) was addeddropwise (an exotherm was observed and the temperature increased to 65°C.). The reaction was stirred at 50° C. for 2 hours, the mixture wasthen cooled to room temperature and poured onto 2 M HCl, stirred for 15minutes and then extracted twice with ethyl acetate. The organic layerswere combined, washed with brine and then saturated aqueous sodiumbicarbonate, dried over magnesium sulfate, filtered and the solventremoved in vacuo to give ethyl5-bromo-1-isopropyl-1H-pyrazole-4-carboxylate (163 g, quantitative)which was used in the next step without further purification. ¹H NMR(400 MHz, CDCl₃, δ): 7.97 (s, 1H), 4.77 (m, 1H), 4.28 (q, 2H), 1.35 (t,3H), 0.90 (d, 6H).

Step 3: (5-bromo-1-isopropyl-1H-pyrazol-4-yl)methanol

To a solution of ethyl 5-bromo-1-isopropyl-1H-pyrazole-4-carboxylate(163 g, 0.50 mol) in 2-methyl tetrahydrofuran (400 mL) was addedborane-DMS (140 mL, 1.50 mol) at 0° C., under argon (effervescenceceased after 50 mL was added). The mixture was stirred at roomtemperature for 30 minutes and then heated to 70° C. for 2 hours, andthen to reflux for 17 hours. Additional portion of borane DMS (40 mL)was added and the mixture was stirred at reflux for an additional 3hours. The mixture was cooled to room temperature then added graduallyto ice-cold methanol (500 mL) with stirring, over a period of 30minutes. The mixture was stirred at room temperature for 30 minutes then2 M aqueous sodium hydroxide (1.5 L) was added. The layers wereseparated and the aqueous layer was extracted with ethyl acetate (2×500mL). The organic layers were combined, washed with brine (500 mL), driedover magnesium sulfate, filtered and the solvent removed in vacuo. Thecrude product was purified by dry flash chromatography (0-50% ethylacetate in heptane) to give(5-bromo-1-isopropyl-1H-pyrazol-4-yl)methanol (70.8 g, 65% over twosteps). +ESI (M+H) 220.9; ¹H NMR (400 MHz, CDCl₃, δ): 7.52 (s, 1H), 4.67(m, 1H), 4.47 (s, 2H), 2.59 (br. s., 1H), 1.41 (s, 6H).

Step 4: 5-bromo-4-(bromomethyl)-1-isopropyl-1H-pyrazole

To a stirred solution of (5-bromo-1-isopropyl-1H-pyrazol-4-yl)methanol(10.0 g, 45.7 mmol) in dichloromethane (200 mL) was added PBr₃ (6.5 mL,68.5 mmol) at 0° C. After the addition was complete the mixture wasallowed to warm to room temperature and stirred for 3 hours. The mixturewas poured into ice-cold water (300 mL), shaken, separated, and thenwashed twice with ice-cold water (2×100 mL) and then brine (100 mL),dried over sodium sulfate, filtered and the solvent removed in vacuo togive 5-bromo-4-(bromomethyl)-1-isopropyl-1H-pyrazole (12.2 g, 95%). ¹HNMR (300 MHz, CDCl₃, δ): 7.58 (s, 1H), 4.64 (m, 1H), 4.35 (s, 2H), 1.43(d, 6H).

Step 5: 1-tert-butyl 4-ethyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate

To a stirred solution of 1-tert-butyl 4-ethylpiperidine-1,4-dicarboxylate (14.5 g, 56.3 mmol) in 2-methyltetrahydrofuran (120 mL) was added, dropwise, 1 M LiHMDS intetrahydrofuran (57 mL, 56.3 mmol) at −78° C. under argon. After 20 min,5-bromo-4-(bromomethyl)-1-isopropyl-1H-pyrazole (12.2 g, 43.3 mmol) in2-methyltetrahydrofuran (10 mL) was added. The mixture was allowed towarm to room temperature and stirred for 18 hours. The mixture wasdiluted with water (200 mL) and the mixture was separated. The organicphase was washed with 10% citric acid solution (2×100 mL), then brine(100 mL), dried over sodium sulfate, filtered and the solvent removed invacuo. The crude product was purified by flash column chromatography(10-30% ethyl acetate in heptane) to give 1-tert-butyl 4-ethyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate(9.3 g). Also isolated from the column was a 7.1 g mixed fraction ofstarting ester and desired product. This was stirred with 1 equivalentof sodium hydroxide in 90% ethanol/methanol for 2 hours at roomtemperature. The solvent was removed in vacuo and ethyl acetate (100 mL)was added. The mixture was washed with 2 N sodium hydroxide (2×50 mL)and then brine (100 mL), dried over sodium sulfate, filtered and thesolvent removed in vacuo to give a second crop of 1-tert-butyl 4-ethyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate(5.1 g). The combined yield is 14.4 g (72%). +ESI (M+H) 404.0; ¹H NMR(400 MHz, CDCl₃, δ): 4.62 (m, 1H), 4.12 (q, 2H), 3.90 (br. s., 2H), 2.82(m, 2H), 2.63 (s, 2H), 2.08 (d, 2H), 1.66 (m, 2H), 1.42 (s, 9H), 1.21(t, 3H).

Step 6:4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylicacid

To a solution of 1-tert-butyl 4-ethyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate(14.5 g, 31.6 mmol) in methanol (50 mL) was added lithium hydroxide(1.52 g, 36.2 mmol) and the mixture was stirred at 80° C. for 18 hours.An additional portion of lithium hydroxide (2.55 g, 63.3 mmol) was addedand the mixture was heated under vigorous reflux for 3 hours, cooled toroom temperature, the solvent was removed in vacuo. The residue waswashed with ethyl acetate, filtered, and the filtrate was saved. Thesolids were dissolved in 2 N aqueous sodium hydroxide (40 mL) and thenacidified to pH 5 with 10% citric acid solution. The aqueous solutionwas extracted with ethyl acetate (3×40 mL), the organics were combined,dried over magnesium sulfate, filtered and then combined with theoriginal filtrate. The solvent was removed from the filtrate underreduced pressure and the resulting residue was purified by flash columnchromatography (ethyl acetate/heptanes) to afford4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylicacid (10.1 g, 74%) as a colorless solid. +ESI (M+H) 429.9; ¹H NMR (300MHz, CDCl₃, δ): 7.41 (s, 1H), 4.64 (m, 1H), 3.94 (m, 2H), 2.95 (m, 2H),2.68 (m, 2H), 2.09 (m, 2H), 1.47 (m, 17H).

Step 7: tert-butyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate

A mixture of4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)-1-(tert-butoxycarbonyl)piperidine-4-carboxylicacid (2.54 g, 5.9 mmol), diphenylphosphoryl azide (1.79 g, 6.5 mmol) andtriethylamine (0.91 mL, 6.5 mmol) in toluene (15 mL) was heated atreflux for 3 hours. The mixture was then cooled to room temperature andthe solvent removed in vacuo. The crude product was purified by columnchromatography to give tert-butyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate(2.8 g, 100%). ¹H NMR (300 MHz, CDCl₃, δ): 7.47 (s, 1H), 4.68 (m, 1H),3.99 (m, 2H), 2.95 (m, 2H), 2.67 (s, 2H), 1.62 (m, 4H), 1.45 (m, 15H).

Step 8: tert-butyl1′-isopropyl-7′-oxo-1′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate

To a mixture of tert-butyl4-((5-bromo-1-isopropyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate(1.4 g, 3.3 mmol) in 2-methyl tetrahydrofuran (10 mL) was added t-butyllithium (1.7 M in hexane, 4.3 mL, 7.2 mmol) at −78° C., under argon.After the addition was complete the mixture was allowed to warm to roomtemperature and was stirred for 18 hours. The mixture was quenched withwater (10 mL) and then diluted with ethyl acetate (20 mL). The layerswere separated and the organic layer was washed with brine (10 mL),dried over sodium sulfate, filtered and the solvent removed in vacuo.The crude product was purified by flash column chromatography to givetert-butyltetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(0.77 g, 67%). +ESI (M+H) 374.1; ¹H NMR (300 MHz, CDCl₃, δ): 7.34 (s,1H), 6.35 (s, 1H), 5.45 (m, 1H), 3.57 (m, 2H), 3.42 (m, 2H), 2.79 (s,2H), 1.70 (m, 4H), 1.45 (m, 15H).

Step 9:1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one

To a solution of tert-butyl1′-isopropyl-7′-oxo-1′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(100 mg, 0.29 mmol) in 4 mL ethyl acetate was added 4 N HCl in dioxane(2 mL). After stirring 30 minutes at room temperature, methanol (1 mL)was added and the resultant solution was stirred for 5 hours at roomtemperature. The volatiles were removed under reduced pressure and theresultant colorless solid triturated with 1:1acetonitrile/dichloromethane to yield 71 mg of the title compound as acolorless solid. ¹H NMR (400 MHz, DMSO-d₆, δ): 8.72 (br. s., 2H), 8.05(s, 1H), 7.37 (s, 1H), 5.36 (m, 1H), 3.15 (m, 2H), 3.05 (m, 2H), 2.78(s, 2H), 1.78 (m, 4H), 1.33 (d, J=6.6 Hz, 6H).

Intermediate 3:2′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt, shown below, was prepared as follows

Step 1: ethyl 3-iodo-1-isopropyl-1H-pyrazole-4-carboxylate

To a solution of ethyl 3-iodopyrazole-4-carboxylate (1.58 g, 5.94 mmol,Truong; et al. Bioorg. Med. Chem. Lett., 19, 4920 (2009)) in 20 mLN,N-dimethylformamide was added cesium carbonate (3.87 g, 11.9 mmol) and2-iodopropane (0.89 mL, 8.90 mmol). The mixture was stirred 2 hours at60° C. and then cooled to ambient temperature. The reaction mixture wasdiluted with 150 mL water and extracted with 2×100 mL diethyl ether. Thecombined organics were washed with 50 mL brine, dried over sodiumsulfate, filtered and concentrated. The resultant oil was purified byflash chromatography (7-60% ethyl acetate/heptane gradient, 50 g silica)to yield 340 mg of ethyl 5-iodo-1-isopropyl-1H-pyrazole-4-carboxylate asa clear oil which crystallized on standing and 740 mg of ethyl3-iodo-1-isopropyl-1H-pyrazole-4-carboxylate as a clear oil. Ethyl5-iodo-1-isopropyl-1H-pyrazole-4-carboxylate: +APCI (M+H) 309.0; ¹H NMR(400 MHz, CDCl₃, δ): 8.05 (s, 1H) 4.82 (spt, J=6.6 Hz, 1H) 4.33 (q,J=7.2 Hz, 2H) 1.50 (d, J=6.6 Hz, 6H) 1.37 (t, J=7.1 Hz, 3H). Ethyl3-iodo-1-isopropyl-1H-pyrazole-4-carboxylate: +APCI (M+H) 309.0; ¹H NMR(400 MHz, CDCl₃, δ): 7.84 (s, 1H) 4.52 (spt, J=6.7 Hz, 1H) 4.32 (q,J=7.1 Hz, 2H) 1.52 (d, J=6.6 Hz, 6H) 1.37 (t, J=7.1 Hz, 3H).

Step 2: (3-iodo-1-isopropyl-1H-pyrazol-4-yl)methanol

A solution of ethyl 3-iodo-1-isopropyl-1H-pyrazole-4-carboxylate (740mg, 2.40 mmol) in tetrahydrofuran (20 mL) was cooled to −78° C. andtreated with diisobutylaluminum hydride (1.5 M in toluene, 0.8 mL, 7.21mmol), dropwise. The mixture was stirred at −78° C. for 1 hour and thenwarmed to room temperature for 2 hours. The mixture was quenched with 10mL ethyl acetate, stirred 15 minutes, and then treated with 25 mLsaturated aqueous Rochelle's salts. After stirring an additional 1 hourat room temperature, the mixture was diluted with 50 mL ethyl acetateand washed with 100 mL water. The aqueous layer was extracted with anadditional 50 mL ethyl acetate. The combined organic layers were driedover sodium sulfate, filtered and concentrated. The residue was thenpurified by flash chromatography (12-100% ethyl acetate/heptanes, 25 gsilica gel) to yield 630 mg of(3-iodo-1-isopropyl-1H-pyrazol-4-yl)methanol as a clear oil. +APCI (M+H)266.8; ¹H NMR (400 MHz, CDCl₃, δ): 7.37 (s, 1H), 4.49 (m, 3H), 1.67 (t,J=5.9 Hz, 1H), 1.50 (s, 6H).

Step 3: 4-(bromomethyl)-3-iodo-1-isopropyl-1H-pyrazole

A solution of (3-iodo-1-isopropyl-1H-pyrazol-4-yl)methanol (0.63 g, 2.37mmol) in 20 mL dichloromethane was cooled to 0° C.Phosphorus(III)bromide (0.67 mL, 7.10 mmol) was added to the solutionand the mixture was stirred 30 minutes at 0° C., 1 hour at roomtemperature, and then quenched with 50 mL water and stirred 15 minutesat room temperature. The mixture was treated with saturated aqueoussodium bicarbonate and extracted with ethyl acetate (2×50 mL). Thecombined organic layers were washed with brine (50 mL), dried oversodium sulfate, filtered and concentrated. The residue was purified byflash chromatography (10-80% ethyl acetate/heptanes, 25 g silica gel) toyield 400 mg of 4-(bromomethyl)-3-iodo-1-isopropyl-1H-pyrazole as acolorless solid. +APCI (M+H) 329.0; ¹H NMR (400 MHz, CDCl₃, δ): 7.42 (s,1H), 4.47 (spt, J=6.7 Hz, 1H), 4.35 (s, 2H), 1.50 (d, J=6.6 Hz, 6H).

Step 4: 1-tert-butyl 4-ethyl4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate

A solution of 1-tert-butyl 4-ethyl piperidine-1,4-dicarboxylate (0.54mL, 2.11 mmol) in tetrahydrofuran (15 mL) in a dry 100 mL round bottomflask under nitrogen was cooled to −78° C. and then treated with lithiumbis(trimethylsilyl) amide (1 M toluene, 2.13 mL, 2.13 mmol). Afterstirring for 45 minutes at −78° C.,4-(bromomethyl)-3-iodo-1-isopropyl-1H-pyrazole (535 mg, 1.63 mmol) wasadded as a suspension in 10 mL tetrahydrofuran. The mixture was stirred1 hour at −78° C. and then allowed to stir 18 hours at room temperature.The reaction mixture was quenched with 20 mL saturated aqueous ammoniumchloride, stirred 30 minutes at room temperature, diluted with 50 mLwater and then extracted with ethyl acetate (2×50 mL). The combinedorganics were dried over sodium sulfate, filtered and concentrated. Theresidue was purified by flash chromatography (10-80% ethylacetate/heptanes, 25 g silica gel) to yield 1-tert-butyl 4-ethyl4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate(645 mg) as a clear oil. +ESI (M-tBu) 450.1; ¹H NMR (400 MHz, CDCl₃, δ):7.02 (s, 3H), 4.44 (spt, J=6.6 Hz, 1H), 4.17 (m, 2H), 3.92 (m, 2H), 2.86(m, 2H), 2.62 (s, 2H), 2.08 (m, 2H), 1.46 (m, 17H), 1.25 (t, J=7.1 Hz,3H).

Step 5:1-(tert-butoxycarbonyl)-4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-4-carboxylicacid

To a solution of 1-tert-butyl 4-ethyl4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-1,4-dicarboxylate(455 mg, 0.9 mmol) in methanol (20 mL) was added 2 N NaOH (5 mL). Afterstirring for 18 hours at room temperature, the methanol was removedunder reduced pressure and the resultant slurry was taken up in 20 mLwater, acidified with 2 N HCl and extracted with ethyl acetate (2×30mL). The combined organic extracts were dried over sodium sulfate,filtered and concentrated to yield1-(tert-butoxycarbonyl)-4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-4-carboxylicacid (430 mg) as a colorless solid. −APCI (M−H) 476.1; ¹H NMR (400 MHz,CDCl₃, δ): 7.11 (s, 1H), 4.45 (dquin, J=13.4, 6.7 Hz, 1H), 3.95 (br. s.,2H), 2.91 (m, 2H), 2.69 (s, 2H), 2.08 (m, 2H), 1.47 (m, 8H).

Step 6: tert-butyl4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate

To a solution of1-(tert-butoxycarbonyl)-4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)piperidine-4-carboxylicacid (430 mg, 0.90 mmol) in toluene (10 mL) was added triethylamine(0.15 mL, 1.08 mmol) and diphenylphosphoryl azide (0.24 mL, 1.08 mmol).The mixture was heated at 120° C. for 3 hours, the volatiles wereremoved under reduced pressure and the resultant oil was purified byflash chromatography (7-60% ethyl acetate/heptanes, 25 g silica gel) toyield tert-butyl4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate(280 mg) as a clear oil. FT-IR (cm⁻¹): 2253; ¹H NMR (400 MHz, CDCl₃, δ):7.27 (s, 1H), 4.50 (m, 1H), 4.03 (br. s., 2H), 2.97 (br. s., 2H), 2.65(s, 2H), 1.65 (m, 4H), 1.50 (s, 6H), 1.47 (s, 9H).

Step 7: tert-butyl2′-isopropyl-7′-oxo-2′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate

To a −78° C. solution of tert-butyl4-((3-iodo-1-isopropyl-1H-pyrazol-4-yl)methyl)-4-isocyanatopiperidine-1-carboxylate(280 mg, 0.59 mmol) in tetrahydrofuran (10 mL) was added t-butyl lithium(0.7 mL, 1.7 M in pentane), dropwise. After stirring for 30 minutes at−78° C. the mixture was warmed to 0° C., quenched with 20 mL saturatedaqueous ammonium chloride, and stirred an additional 30 minutes at roomtemperature. The reaction mixture was diluted with 25 mL water andextracted with ethyl acetate (2×50 mL). The combined organics were driedover sodium sulfate, filtered and concentrated. The residue was thenpurified by flash chromatography (12-100% ethyl acetate/heptanes, 10 gsilica gel) to yield tert-butyl2′-isopropyl-7′-oxo-2′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(130 mg) as a colorless solid. +ESI (M+H) 349.1; ¹H NMR (500 MHz, CDCl₃,δ): 7.28 (s, 1H), 5.78 (s, 1H), 4.57 (spt, J=6.6 Hz, 1H), 3.59 (m, 2H),3.37 (m, 2H), 2.82 (s, 2H), 1.74 (m, 4H), 1.55 (d, J=6.6 Hz, 6H), 1.47(s, 9H).

Step 8:2′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride

To a solution of tert-butyl2′-isopropyl-7′-oxo-2′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(130 mg, 0.37 mmol) in ethyl acetate (5 mL) was added 4 M hydrochloricacid (2 mL) in 1,4-dioxane. The reaction mixture was stirred 3 hours atroom temperature, the volatiles were removed under reduced pressure andthe resultant residue was triturated with 10 mL heptane. The solid wasdried under reduced pressure to yield2′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride (105 mg) as an off-white solid. +ESI (M+H) 249.1; ¹H NMR(500 MHz, DMSO-d₆, δ): 7.91 (s, 1H) 7.69 (s, 1H) 4.48-4.62 (m, 1H)3.02-3.28 (m, 4H) 2.78 (s, 2H) 1.74-1.89 (m, 4H) 1.41 (d, J=6.59 Hz,6H).

Intermediate 4:2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt, shown below, was prepared as follows

Step 1: (E)-ethyl 2-(2-tert-butylhydrazono)propanoate

To a solution of ethyl pyruvate (20.22 g, 174.1 mmol) in ethanol (150mL) was added t-butyl hydrazine hydrochloride (21.7 g, 174 mmol) andN,N-diisopropylethyl amine (33.4 mL, 192 mmol). After stirring at refluxfor 18 hours, the reaction was cooled and the volatiles were removedunder reduced pressure. The resultant golden oil was taken up in 300 mLethyl acetate and washed with 200 mL water and 300 mL saturated aqueoussodium bicarbonate. The organic layer was dried over sodium sulfate,filtered and concentrated to yield (E)-ethyl2-(2-tert-butylhydrazono)propanoate (23.1 g) as a clear pale yellow oil.+APCI (M+H) 187.3; ¹H NMR (400 MHz, CDCl₃, δ): 5.51 (br. s., 1H), 4.25(q, J=7.2 Hz, 2H), 1.89 (s, 3H), 1.32 (t, J=7.1 Hz, 3H), 1.28 (s, 9H).

Step 2: ethyl 1-tert-butyl-4-formyl-1H-pyrazole-3-carboxylate

To a yellow orange solution of (E)-ethyl2-(2-tert-butylhydrazono)propanoate (22.9 g, 123 mmol) in toluene (300mL) was added (chloromethylene)dimethylammonium chloride (Vilsmeiersalt, 34.0 g, 252 mmol) in a single portion. The suspension was stirred3 hours at room temperature, slowly becoming a biphasic mixture oftoluene over a thick orange oil. The reaction mixture was cooled to 0°C. and slowly neutralized with saturated aqueous sodium bicarbonate. Thelayers were separated and the aqueous layer extracted with additionalethyl acetate (2×200 mL). The organic layers were combined, washed with200 mL brine, dried over sodium sulfate, filtered and concentrated toyield ethyl 1-tert-butyl-4-formyl-1H-pyrazole-3-carboxylate (18.6 g) asa tan-orange oil which solidified on standing. +APCI (M+H) 225.1; ¹H NMR(400 MHz, CDCl₃, δ): 10.37 (s, 1H), 8.14 (s, 1H), 4.48 (q, J=7.0 Hz,2H), 1.65 (s, 9H), 1.44 (t, 3H).

Step 3: ethyl 1-tert-butyl-4-(hydroxymethyl)-1H-pyrazole-3-carboxylate

To a solution of ethyl 1-tert-butyl-4-formyl-1H-pyrazole-3-carboxylate(2.87 g, 12.8 mmol) in ethanol (50 mL) was added sodium borohydride(0.97 g, 25.6 mmol) in one portion. After stirring for 30 minutes atroom temperature the mixture was quenched with 1 N aqueous hydrochloricacid (100 mL), stirred for 15 minutes, and then neutralized withsaturated aqueous sodium bicarbonate. The mixture was extracted withethyl acetate (2×150 mL), the combined organics then dried over sodiumsulfate, filtered and concentrated to yield ethyl1-tert-butyl-4-(hydroxymethyl)-1H-pyrazole-3-carboxylate (2.57 g) as aclear oil. +APCI (M+Na) 249.2; ¹H NMR (400 MHz, CDCl₃, δ): 7.49 (s, 1H),4.65 (d, J=6.8 Hz, 2H), 4.43 (q, J=7.2 Hz, 2H), 3.62 (t, J=6.9 Hz, 1H),1.59 (s, 9H), 1.41 (t, J=7.1 Hz, 3H).

Step 4: ethyl 4-(bromomethyl)-1-tert-butyl-1H-pyrazole-3-carboxylate

To a 0° C. solution of ethyl1-tert-butyl-4-(hydroxymethyl)-1H-pyrazole-3-carboxylate (3.9 g, 17.24mmol) in dichloromethane (120 mL) was added phosphorus tribromide (4.91mL, 51.7 mmol), and the resultant mixture was stirred 30 minutes at 0°C. and then 1 hour at room temperature. The mixture was quenched with 50mL water, neutralized with saturated aqueous sodium bicarbonate, stirred30 minutes, and then extracted with dichloromethane (2×150 mL). Thecombined organic extracts were washed with 100 mL brine, dried oversodium sulfate, filtered and concentrated. The resulting residue waspurified by flash chromatography (7-60% ethyl acetate/heptanes, 50 gsilica gel) to yield ethyl4-(bromomethyl)-1-tert-butyl-1H-pyrazole-3-carboxylate (4.12 g) as aclear oil. +APCI (M+H) 289.1; ¹H NMR (400 MHz, CDCl₃, δ): 7.61 (s, 1H),4.70 (s, 2H), 4.41 (q, J=7.2 Hz, 2H), 1.60 (s, 9H), 1.40 (t, 3H).

Step 5: tert-butyl4-((1-tert-butyl-3-(ethoxycarbonyl)-1H-pyrazol-4-yl)methyl)-4-cyanopiperidine-1-carboxylate

To a −78° C. solution of tert-butyl4-cyanopiperidine-1-carboxylatepiperidine (1.0 g, 4.76 mmol) intetrahydrofuran (20 mL) was added lithium bis(trimethylsilyl)amide (4.76mL, 1 M in tetrahydrofuran). The mixture was stirred 30 minutes at −78°C., warmed to 0° C. for 30 minutes and then cooled to −78° C. A solutionof ethyl 4-(bromomethyl)-1-tert-butyl-1H-pyrazole-3-carboxylate (1.38 g,4.76 mmol) in tetrahydrofuran was then added, dropwise. After stirring30 minutes at −78° C. the mixture was allowed to warm to roomtemperature and stir an additional 18 hours. The reaction mixture wasquenched with saturated aqueous ammonium chloride (50 mL), stirred for30 minutes, diluted with water (50 mL) and then extracted with ethylacetate (2×50 mL). The organics were combined, dried over sodiumsulfate, filtered and concentrated. The residue was purified by flashchromatography (7-60% ethyl acetate/heptanes, 100 g silica gel) to yieldtert-butyl4-((1-tert-butyl-3-(ethoxycarbonyl)-1H-pyrazol-4-yl)methyl)-4-cyanopiperidine-1-carboxylate(455 mg) as a clear oil. +APCI (M+H) 419.3; ¹H NMR (500 MHz, CDCl₃, δ):7.68 (s, 1H), 4.40 (q, J=7.2 Hz, 2H), 4.13 (br. s., 2H), 3.17 (s, 2H),2.97 (br. s., 2H), 1.81 (d, J=13.2 Hz, 2H), 1.63 (s, 9H), 1.56 (m, 2H),1.46 (s, 9H), 1.41 (t, J=7.2 Hz, 3H).

Step 6:4-((1-(tert-butoxycarbonyl)-4-carbamoylpiperidin-4-yl)methyl)-1-tert-butyl-1H-pyrazole-3-carboxylicacid

To a 0° C. solution of tert-butyl4-((1-tert-butyl-3-(ethoxycarbonyl)-1H-pyrazol-4-yl)methyl)-4-cyanopiperidine-1-carboxylate(455 mg, 1.09 mmol) in methanol (11 mL) was added a solution ofurea-hydrogen peroxide (1.05 g, 10.9 mmol) in 1 M aqueous sodiumhydroxide (10.9 mL), dropwise. After stirring for 18 hours at roomtemperature, volatiles were removed under reduced pressure and theresultant slurry was taken up in water (50 mL), acidified with 2 Naqueous hydrochloric acid and extracted with ethyl acetate (2×50 mL).The combined organic extracts were dried over sodium sulfate, filteredand concentrated to yield4-((1-(tert-butoxycarbonyl)-4-carbamoylpiperidin-4-yl)methyl)-1-tert-butyl-1H-pyrazole-3-carboxylicacid (418 mg) as a colorless solid. −APCI (M−H) 407.3; ¹H NMR (400 MHz,DMSO-d₆, δ): 12.33 (br. s., 1H), 7.47 (s, 1H), 7.11 (br. s., 1H), 6.99(s, 1H), 3.59 (d, J=13.3 Hz, 2H), 2.89 (s, 2H), 2.77 (m, 2H), 1.84 (m,2H), 1.44 (s, 9H), 1.31 (s, 9H), 1.16 (m, 2H).

Step 7:4-((1-(tert-butoxycarbonyl)-4-isocyanatopiperidin-4-yl)methyl)-1-tert-butyl-1H-pyrazole-3-carboxylicacid

To a suspension of4-((1-(tert-butoxycarbonyl)-4-carbamoylpiperidin-4-yl)methyl)-1-tert-butyl-1H-pyrazole-3-carboxylicacid (388 mg, 0.95 mmol) in acetonitrile (20 mL) was added sodiumbicarbonate (319 mg, 3.80 mmol) and bis(trifluoroacetoxy) iodosobenzene(632 mg, 1.42 mmol). The mixture was stirred 90 minutes at roomtemperature, diluted with 50 mL water, acidified with 1 N aqueoushydrochloric acid, and then extracted with ethyl acetate (2×50 mL). Thecombined organic extracts were dried over sodium sulfate, filtered andconcentrated. The resulting residue was purified by flash chromatography(1-10% methanol/dichloromethane, 25 g silica gel) to yield4-((1-(tert-butoxycarbonyl)-4-isocyanatopiperidin-4-yl)methyl)-1-tert-butyl-1H-pyrazole-3-carboxylicacid (172 mg) as a colorless solid. −APCI (M−H) 405.4; ¹H NMR (400 MHz,DMSO-d₆, δ): 12.52 (br. s., 1H), 7.82 (s, 1H), 3.83 (br. s., 2H), 3.03(s, 2H), 2.82 (br. s., 2H), 1.49 (m, 13H), 1.36 (m, 9H).

Step 8: tert-butyl2′-tert-butyl-7′-oxo-2′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate

A solution of4-((1-(tert-butoxycarbonyl)-4-isocyanatopiperidin-4-yl)methyl)-1-tert-butyl-1H-pyrazole-3-carboxylicacid (180 mg, 0.44 mmol) in tetrahydrofuran (5 mL) was treated with 2 Naqueous sodium hydroxide (0.664 mL, 1.33 mmol). The mixture was stirred3 hours at room temperature, tetrahydrofuran and water were removed on arotary evaporator and the resultant colorless solid was slurried inacetonitrile (10 mL) and then concentrated to dryness. The triturationwas repeated twice more from acetonitrile (10 mL). The resultantcolorless solid was taken up in dichloromethane (10 mL) and treated with(3-(dimethylamino)propyl)ethyl carbodiimide hydrochloride (170 mg, 0.89mmol). The mixture was stirred 18 hours at room temperature and thendiluted with dichloromethane (50 mL) and washed with water (30 mL). Theorganic phase was dried over sodium sulfate, filtered and concentrated.The residue was then purified by flash chromatography (30-100% ethylacetate/heptanes, 10 g silica gel) to yield tert-butyl2′-tert-butyl-7′-oxo-2′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(70 mg) as a colorless solid. +ESI (M+H) 363.3; ¹H NMR (400 MHz,DMSO-d₆, δ): 7.68 (s, 1H), 7.57 (s, 1H), 3.47 (m, 2H), 3.20 (m, 2H),2.73 (s, 2H), 1.53 (t, J=5.7 Hz, 4H), 1.49 (s, 9H), 1.36 (s, 9H).

Step 9: The title compound,2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt

To a solution of tert-butyl2′-tert-butyl-7′-oxo-2′,4′,6′,7′-tetrahydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-1-carboxylate(70 mg, 0.19 mmol) in ethyl acetate (5 mL) was added 4 M hydrochloricacid in 1,4-dioxane (2 mL) and the mixture was stirred 3 hours at roomtemperature. The volatiles were removed under reduced pressure and theresultant colorless solid was triturated from heptanes (10 mL) and driedunder reduced pressure to yield2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt (56 mg) as an off-white solid. +ESI (M+H) 263.1; ¹HNMR (500 MHz, DMSO-d₆, δ): 8.72 (m, 2H), 7.92 (s, 1H), 7.75 (s, 1H),3.20 (br. s, 2H), 3.09 (br. s., 2H), 2.78 (s, 2H), 1.79 (m, 4H), 1.48(s, 9H).

Intermediate 5:2′-tert-pentyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one,shown below, was prepared as follows

Step 1: ethyl 3-bromo-1H-pyrazole-4-carboxylate

To a 0° C. solution of ethyl 3-amino-1H-pyrazole-4-carboxylate (5.0 g,32 mmol) and copper (II) bromide (7.2 g, 32 mmol) in acetonitrile (65mL) was slowly added isoamyl nitrite (12 mL, 86 mmol). The reaction washeated to 50° C. and stirred overnight. The reaction was cooled to roomtemperature and quenched with 1 N aqueous hydrochloric acid (150 mL).The mixture was extracted with ethyl acetate (3×100 mL). The combinedorganics were washed with water, dried over sodium sulfate, filtered,and concentrated to give the title compound as a brown oil thatpartially solidified under vacuum overnight (7.1 g, 100%). ¹H NMR (400MHz, CDCl₃, δ): 9.78 (br. s., 1H), 8.10 (br. s., 1H), 4.33 (q, J=7.22Hz, 2H), 1.36 (m, 3H).

Step 2: (3-bromo-1-tert-pentyl-1H-pyrazol-4-yl)methanol

Concentrated sulfuric acid (0.45 mL, 4.8 mmol) was added to a mixture ofethyl 3-bromo-1H-pyrazole-4-carboxylate (1.0 g, 4.6 mmol) and tert-amylalcohol (3.0 mL, 27 mmol). The reaction was heated to 100° C. for 2.5hours. The reaction was then cooled to room temperature and leftstirring overnight. The reaction was diluted with ethyl acetate andwashed with water. The organic layer was dried over sodium sulfate,filtered, and concentrated to yield ethyl3-bromo-1-tert-pentyl-1H-pyrazole-4-carboxylate (1.3 g) as a crude brownoil.

This crude product (1.3 g) was dissolved in tetrahydrofuran (24 mL) andcooled to −78° C. A solution of diisobutylaluminum hydride (1.5 M intoluene, 9.0 mL, 160 mmol) was slowly added, and the reaction wasstirred at −78° C. for 1 hour. The reaction was then allowed to warm toroom temperature and stir for another 2 hours. The reaction was dilutedwith ethyl acetate (20 mL) and saturated aqueous Rochelle's salt (20mL).

The mixture was stirred at room temperature overnight. The layers wereseparated and the aqueous layer was extracted with ethyl acetate. Thecombined organics were washed with brine, dried over sodium sulfate,filtered, and concentrated. Purification by flash column chromatography(0-100% ethyl acetate/heptanes) gave the title compound (685 mg, 62%) asa pale yellow oil. ¹H NMR (400 MHz, CDCl₃, δ): 7.45 (s, 1H), 4.51 (s,2H), 1.86 (q, J=7.41 Hz, 2H), 1.66 (s, 1H), 1.51 (s, 6H), 0.69 (m, 3H).

Step 3: 3-bromo-4-(chloromethyl)-1-tert-pentyl-1H-pyrazole

A solution of (3-bromo-1-tert-pentyl-1H-pyrazol-4-yl)methanol (675 mg,2.73 mmol) in dichloromethane (10 mL) was cooled to 0° C. Triethylamine(0.53 mL, 3.8 mmol) and methanesulfonyl chloride (0.28 mL, 3.6 mmol)were added. The reaction was stirred at 0° C. for 15 minutes, thenwarmed to room temperature and stirred for 1.5 hours. The reaction wasdiluted with ethyl acetate, washed with water and brine, dried oversodium sulfate, filtered, and concentrated to give the title compound(725 mg, 100%) as a clear oil. ¹H NMR (400 MHz, CDCl₃, δ): 7.48 (s, 1H),4.47 (s, 2H), 1.86 (q, J=7.48 Hz, 2H), 1.52 (s, 6H), 0.69 (m, 3H).

Step 4: 3-bromo-4-(iodomethyl)-1-tert-pentyl-1H-pyrazole

To a solution of 3-bromo-4-(chloromethyl)-1-tert-pentyl-1H-pyrazole (725mg, 2.73 mmol) in acetone (25 mL) was added sodium iodide (4.09 g, 27.3mmol). The reaction was heated at reflux for 2 hours, then cooled toroom temperature and stirred overnight. The solvent was removed in vacuoand the residue was partitioned between ethyl acetate and water. Theorganic layer was washed with saturated aqueous sodium thiosulfate andbrine. The organics were dried over sodium sulfate, filtered, andconcentrated to yield the title compound (824 mg, 85%) as a brown oil.¹H NMR (400 MHz, CDCl₃, δ): 7.47 (s, 1H), 4.26 (s, 2H), 1.83 (q, J=7.41Hz, 2H), 1.50 (s, 6H), 0.67 (t, J=7.51 Hz, 3H).

Step 5:2′-tert-pentyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

The title compound was prepared by a method analogous to that describedfor Intermediate 3 in Steps 4-8, using3-bromo-4-(iodomethyl)-1-tert-pentyl-1H-pyrazole. +ESI (M+H) 277.3; ¹HNMR (400 MHz, CD₃OD, δ): 7.67 (s, 1H), 3.22-3.37 (m, 4H), 2.93 (s, 2H),1.92 (q, J=7.61 Hz, 2H), 1.88-2.05 (m, 4H), 1.57 (s, 6H), 0.67 (t,J=7.41 Hz, 3H).

Intermediate 6:2′-cyclobutyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one,shown below, was prepared as follows

Step 1: ethyl 3-bromo-1-cyclobutyl-1H-pyrazole-4-carboxylate

A mixture of ethyl 3-bromo-1H-pyrazole-4-carboxylate (1.00 g, 4.56mmol), cyclobutyl bromide (0.65 mL, 6.9 mmol), and cesium carbonate(2.97 g, 9.13 mmol) in N,N-dimethylformamide (10 mL) was heated to 60°C. and stirred overnight. The reaction was cooled to room temperatureand partitioned between 1:1 heptanes/ethyl acetate and water. Theaqueous was extracted again with 1:1 heptanes/ethyl acetate. Thecombined organics were washed with brine, dried over sodium sulfate,filtered, and concentrated. Purification by flash column chromatographygave two product regioisomers as colorless oils.

ethyl 5-bromo-1-cyclobutyl-1H-pyrazole-4-carboxylate (230 mg, 18%): +ESI(M+H+1) 275.1; ¹H NMR (400 MHz, CDCl₃, δ): 7.98 (s, 1H), 4.98 (m, 1H),4.30 (q, J=7.02 Hz, 2H), 2.61-2.74 (m, 2H), 2.43 (m, 2H), 1.84-1.95 (m,2H), 1.34 (m, 3H).

ethyl 3-bromo-1-cyclobutyl-1H-pyrazole-4-carboxylate (570 mg, 46%): +ESI(M+H+1) 275.1; ¹H NMR (400 MHz, CDCl₃, δ): 7.87 (s, 1H), 4.69 (m, 1H),4.29 (q, J=7.22 Hz, 2H), 2.41-2.61 (m, 4H), 1.78-1.98 (m, 2H), 1.34 (m,3H).

Step 2: (3-bromo-1-cyclobutyl-1H-pyrazol-4-yl)methanol

A solution of ethyl 3-bromo-1-cyclobutyl-1H-pyrazole-4-carboxylate (565mg, 2.07 mmol) in tetrahydrofuran (10 mL) was cooled to −78° C.Diisobutylaluminum hydride (4.13 mL, 6.02 mmol, 1.5 M in toluene) wasadded slowly and the reaction was stirred at −78° C. for 1 hour. Thereaction was then allowed to warm to room temperature and stir for anadditional 2 hours. The reaction was diluted with ethyl acetate (20 mL)and saturated aqueous Rochelle's salt (20 mL). The mixture was stirredat room temperature overnight. The mixture was further diluted withethyl acetate and was washed with water. The aqueous layer was extractedwith ethyl acetate and the combined organics were dried over sodiumsulfate, filtered, and concentrated to yield the title compound (478 mg,100%) as a colorless oil. +APCI (M+H+1) 233.1; ¹H NMR (400 MHz, CDCl₃,δ): 7.40 (s, 1H), 4.62-4.71 (m, 1H), 4.51 (s, 2H), 2.39-2.59 (m, 4H),1.74-1.92 (m, 3H).

Step 3:2′-cyclobutyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

The title compound was prepared by a method analogous to that describedfor Intermediate 5, Steps 3-5, using(3-bromo-1-cyclobutyl-1H-pyrazol-4-yl)methanol. +ESI (M+H) 261.3; ¹H NMR(400 MHz, CD₃OD, 6): 7.62 (s, 1H), 4.84-4.92 (m, 1H), 3.21-3.36 (m, 4H),2.93 (s, 2H), 2.50-2.63 (m, 2H), 2.40-2.50 (m, 2H), 1.82-2.05 (m, 6H).

Intermediate 7:2′-tert-butyl-4′,6′-dihydro-8-azaspiro[bicyclo[3.2.1]octane-3,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride, shown below, was prepared as follows

Step 1: ethyl 3-iodo-1H-pyrazole-4-carboxylate

Ethyl 3-amino-1H-pyrazole-4-carboxylate (860.0 mg, 5.54 mmol) wasdissolved in acetic acid (5 mL) and water (5 mL). Potassium iodide (921mg, 5.54 mmol) was added and the mixture was stirred until all solidshad dissolved. A solution of sodium nitrite (386 mg, 5.54 mmol) in water(2 mL) was then added dropwise. The reaction was stirred at roomtemperature for 2 minutes when stirring became hindered due toprecipitate formation. Additional water (5 mL) was added and thereaction was allowed to stir overnight. The acetic acid was removedunder reduced pressure. The brown residue was taken up in saturatedaqueous sodium bicarbonate and was extracted with ethyl acetate (2×50mL). The combined organics were washed with saturated aqueous sodiumthiosulfate (50 mL), dried over sodium sulfate, filtered, andconcentrated. Purification by flash column chromatography (10-80% ethylacetate/heptanes) gave the title compound (863 mg, 59%) as a whitesolid. +APCI (M+H) 267.2; ¹H NMR (400 MHz, CDCl₃, δ): 12.63 (br. s.,1H), 8.13 (s, 1H), 4.34 (q, J=7.0 Hz, 2H), 1.38 (t, J=7.2 Hz, 3H).

Step 2: ethyl 1-tert-butyl-3-iodo-1H-pyrazole-4-carboxylate

To a solution of ethyl 3-iodo-1H-pyrazole-4-carboxylate (1.10 g, 3.91mmol) in tert-butanol (5 mL) was added sulfuric acid (0.40 mL, 4.18mmol, 18 M). The reaction was heated to 100° C. and stirred for 3 hours.The reaction was cooled to room temperature and diluted with ethylacetate (100 mL) and water (25 mL). The pH was adjusted to 8 withsaturated aqueous sodium bicarbonate. The layers were separated and theorganics were dried over sodium sulfate, filtered, and concentrated.Purification by flash column chromatography (7-60% ethylacetate/heptanes) gave 2 regioisomeric products.

ethyl 1-tert-butyl-5-iodo-1H-pyrazole-4-carboxylate: ¹H NMR (400 MHz,CDCl₃, δ): 7.91 (s, 1H), 4.32 (q, J=7.2 Hz, 2H), 1.83 (s, 9H), 1.37 (t,J=7.1 Hz, 3H).

ethyl 1-tert-butyl-3-iodo-1H-pyrazole-4-carboxylate (976 mg, 73%) as aclear oil: +APCI (M+H) 323.3; ¹H NMR (400 MHz, CDCl₃, δ): 7.90 (s, 1H),4.31 (q, J=7.0 Hz, 2H), 1.59 (s, 9H), 1.36 (t, J=7.1 Hz, 3H).

Step 3: 1-tert-butyl-3-iodo-4-(iodomethyl)-1H-pyrazole

The title compound was prepared by a method analogous to that describedfor Intermediate 5, Steps 2-4, using ethyl1-tert-butyl-3-iodo-1H-pyrazole-4-carboxylate. ¹H NMR (400 MHz, CDCl₃,δ): 7.49 (s, 1H), 4.25 (s, 2H), 1.56 (s, 9H).

Step 4: (1R,5S)-8-tert-butyl 3-methyl8-azabicyclo[3.2.1]octane-3,8-dicarboxylate

To a solution of(1R,5S)-8-(tert-butoxycarbonyl)-8-azabicyclo[3.2.1]octane-3-carboxylicacid (500 mg, 1.96 mmol) in N,N-dimethylformamide (5 mL) was addedpotassium carbonate (541 mg, 3.92 mmol) and methyl iodide (0.18 mL, 2.94mmol). The reaction was stirred at room temperature for 18 hours. Thereaction was diluted with ethyl acetate (50 mL) and heptanes (50 mL),and then washed with water (100 mL) and brine (50 mL). The organics weredried over sodium sulfate, filtered, and concentrated. Purification byflash column chromatography gave the title compound (486 mg, 92%) as aclear oil. ¹H NMR (400 MHz, CDCl₃, δ): 4.17-4.29 (m, 2H), 3.65 (s, 3H),2.75-2.86 (m, 1H), 1.93-2.01 (m, 2H), 1.79-1.92 (m, 2H), 1.67-1.76 (m,2H), 1.58-1.66 (m, 2H), 1.45 (s, 9H).

Step 5:2′-tert-butyl-4′,6′-dihydro-8-azaspiro[bicyclo[3.2.1]octane-3,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride

The title compound was prepared by a method analogous to that describedfor Intermediate 3, Steps 4-8, using (1R,5S)-8-tert-butyl 3-methyl8-azabicyclo[3.2.1]octane-3,8-dicarboxylate and1-tert-butyl-3-iodo-4-(iodomethyl)-1H-pyrazole. +ESI (M+H) 289.2; ¹H NMR(400 MHz, CD₃OD, δ): 7.69 (s, 1H), 4.03-4.10 (m, 2H), 2.74 (s, 2H),2.39-2.46 (m, 2H), 2.10-2.25 (m, 6H), 1.59 (s, 9H).

Intermediate 8: 1 H-pyrazolo[4,3-b]pyridine-6-carboxylic acid, shownbelow, was prepared as follows:

Step 1: diethyl 2-(5-bromo-3-nitropyridin-2-yl)malonate

To a suspension of sodium hydride (5.08 g, 127 mmol) inN,N-dimethylformamide (75 mL) was added diethyl malonate (19.26 mL, 127mmol) at 0° C. The solution was then stirred at ambient temperature for30 minutes and a solution of 5-bromo-2-chloro-3-nitropyridine (30 g, 127mmol) in N,N-dimethylformamide (75 mL) was added dropwise. The darkbrown mixture was then stirred at 100° C. for 2 hours before beingcooled to ambient temperature and quenched with a saturated solution ofammonium chloride (500 mL) at 0° C. The mixture was extracted with ethylacetate (3×500 mL) and the combined organics were dried over magnesiumsulfate and filtered. The solvent was removed in vacuo to give a darkbrown oil which was purified by flash column chromatography (10% ethylacetate/hexane) to afford diethyl2-(5-bromo-3-nitropyridin-2-yl)malonate as a brown solid (31.8 g, 69%).¹HNMR (400 MHz, CDCl₃, δ): 8.86 (s, 1H), 8.61 (s, 1H), 5.44 (s, 1H),4.29 (q, 4H), 1.27 (t, 6H).

Step 2: 5-bromo-2-methyl-3-nitropyridine

A mixture of the diethyl 2-(5-bromo-3-nitropyridin-2-yl)malonate (31.8g, 88 mmol) in aqueous hydrochloric acid (6 M, 1.4 L) was stirred atreflux for 18 hours. The reaction mixture was cooled to ambienttemperature and added very slowly to a saturated aqueous solution ofaqueous sodium bicarbonate (4 L) at 0° C. The mixture was then extractedwith dichloromethane (7 L), dried over magnesium sulfate and the solventremoved in vacuo to give 5-bromo-2-methyl-3-nitropyridine as an orangeoil (13.8 g, 72%) which solidified upon standing. ¹HNMR (300 MHz, CDCl₃,δ): 8.78 (s, 1H), 8.43 (s, 1H), 2.79 (s, 3H).

Step 3: 5-bromo-2-methylpyridin-3-amine

To a solution of 5-bromo-2-methyl-3-nitropyridine (13.8 g, 63.9 mmol) inindustrial methylated spirit (330 mL) at 40° C. was added iron powder(20 g) (portionwise to avoid clumping) followed by concentrated aqueoushydrochloric acid (5 mL). The dark brown mixture was stirred vigorouslyat reflux for 2 hours and then cooled and filtered through Celite®(which was washed with 1 L of industrial methylated spirit). The solventwas then removed in vacuo and the residue taken up in ethyl acetate (200mL) and washed with a saturated aqueous solution of sodium bicarbonate(200 mL), dried over magnesium sulfate and solvent removed in vacuo togive 5-bromo-2-methylpyridin-3-amine as an orange solid (10.7 g, 90%).¹HNMR (400 MHz, CDCl₃, δ): 7.91 (s, 1H), 7.00 (s, 1H), 3.75 (br.s., 2H),2.25 (s, 3H).

Step 4: N-(5-bromo-2-methylpyridin-3-yl)acetamide

To a solution of 5-bromo-2-methylpyridin-3-amine (10.7 g, 57.5 mmol) indichloromethane (575 mL) was added acetic anhydride (12 mL, 126.5 mmol)at 0° C. followed by triethylamine (22 mL, 158 mmol). The mixture wasallowed to warm to ambient temperature and stirred for 18 hours at whichpoint a further equivalent of acetic anhydride (6 mL, 63 mmol) wasadded. The mixture was stirred at ambient temperature for a further 72hours. The reaction mixture was quenched with a saturated aqueoussolution of sodium bicarbonate (500 mL) and the organic phase washedwith saturated aqueous sodium chloride (500 mL), dried over magnesiumsulfate and concentrated in vacuo to give a brown solid. This solid wastriturated with 30% ethyl acetate in hexanes to yieldN-(5-bromo-2-methylpyridin-3-yl)acetamide as an off-white solid (8.28 g,63%). ¹HNMR (400 MHz, CD₃OD, δ): 8.31 (s, 1H), 8.18 (s, 1H), 2.43 (s,3H), 2.18 (s, 3H).

Step 5: 6-bromo-1H-pyrazolo[4,3-b]pyridine

To a solution of N-(5-bromo-2-methylpyridin-3-yl)acetamide (8.28 g, 36mmol) in chloroform (550 mL) at ambient temperature was added potassiumacetate (4.32 g, 43.6 mmol), acetic acid (2.5 mL, 43.6 mmol) andfollowed by acetic anhydride (6.86 mL, 72.6 mmol). The mixture wasstirred at ambient temperature for 15 minutes before being heated to 40°C. Isoamylnitrite was then added dropwise. The reaction was then stirredat 60° C. for 48 hours. The reaction mixture was poured slowly into asaturated solution of sodium bicarbonate (500 mL) at 0° C. The organicphase was retained and the aqueous phase extracted with dichloromethane(500 mL). The combined organics were then concentrated to a brown oilwhich was dissolved in methanol (500 mL). Aqueous sodium hydroxide (2 M,500 mL) was added at 0° C. and the mixture stirred at ambienttemperature for 1 hour before the methanol was removed in vacuo. Theaqueous mixture was then extracted with ethyl acetate (3×500 mL). Thecombined organics were dried over magnesium sulfate, filtered, and thesolvent removed in vacuo to give 6-bromo-1H-pyrazolo[4,3-b]pyridine as alight brown solid (5.5 g, 77%). ¹HNMR (400 MHz, CD₃OD, δ): 8.55 (s, 1H),8.24 (s, 1H), 8.21 (s, 1H).

Step 6: methyl 1H-pyrazolo[4,3-b]pyridine-6-carboxylate

To a solution of 6-bromo-1H-pyrazolo[4,3-b]pyridine (5.5 g, 27.9 mmol)in methanol (125 mL) and acetonitrile (75 mL) was added triethylamine(22 mL, 156 mmol), 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (1.98 g,3.07 mmol), and palladium dichloride (1.23 g, 6.98 mmol). The mixturewas placed under 20 bar of carbon monoxide, heated to 100° C., andstirred vigorously for 18 hours. The reaction mixture was cooled toambient temperature and filtered through Celite® before the solvent wasremoved in vacuo to yield a brown oil. This crude oil was then purifiedby flash column chromatography (50% ethyl acetate/hexanes) to givemethyl 1H-pyrazolo[4,3-b]pyridine-6-carboxylate as a pale yellow solid(4.52 g, 92%). ¹HNMR (400 MHz, CDCl₃, δ): 10.56 (s, 1H), 9.23 (s, 1H),8.35 (s, 1H), 8.40 (s, 1H), 4.01 (s, 3H).

Step 7: 1H-pyrazolo[4,3-b]pyridine-6-carboxylic acid

To a solution of methyl 1H-pyrazolo[4,3-b]pyridine-6-carboxylate (3.52g, 20 mmol) in methanol (250 mL) and water (190 mL) at 0° C. was addedaqueous sodium hydroxide (2M, 64 mL, 128 mmol), dropwise. The suspensionwas then allowed to warm to ambient temperature and stirred for 18hours. The methanol was then removed in vacuo and the aqueous mixtureextracted with ethyl acetate (250 mL). The aqueous layer was acidified(to pH 5-6) with 2 N aqueous hydrochloric acid (70 mL). The cream solidwhich had precipitated out was then filtered off and dried in adesiccator to yield the title compound (0.675 g, 21%). ¹HNMR (400 MHz,DMSO-d₆, δ): 8.97 (s, 1H), 8.45 (s, 1H), 8.39 (s, 1H).

Intermediate 9: 3-carbamoyl-1H-indazole-5-carboxylic acid, shown below,was prepared as follows

Step 1. methyl 3-cyano-1H-indazole-5-carboxylate

Methyl 3-iodo-1H-indazole-5-carboxylate (30.7 g, 102 mmol), zinc cyanide(20.3 g, 173 mmol), zinc dust (4.05 g, 61.9 mmol),[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II), complexwith dichloromethane (12 g, 15 mmol), and copper (I) iodide (19.7 g, 103mmol) were combined in a 1 L round bottom flask. N,N-dimethylacetamide(500 mL) was added and the reaction mixture was purged with nitrogen for10 minutes. The reaction was heated to 120° C. for 1 hour. The reactionwas cooled to room temperature and was diluted with ethyl acetate (1 L),and allowed to stir for 20 minutes. The reaction mixture was filteredthrough a plug of Celite, rinsing with 500 mL ethyl acetate. Thefiltrate was added to a solution of saturated ammonium chloride andconcentrated ammonium hydroxide (2 L) (prepared by adding ammoniumhydroxide to a saturated aqueous solution of ammonium chloride untilpH=8) and the biphasic solution was stirred vigorously for 1 hour. Theresulting emulsion was filtered through a small pad of Celite. Thelayers were separated and the aqueous was extracted two additional timeswith ethyl acetate (1100 mL), each time filtering the resulting emulsionthrough Celite. The combined organic layers were washed with water(2×900 mL) and brine (900 mL), dried over sodium sulfate, filtered andconcentrated. To the crude was added methanol (100 mL) and the mixturewas stirred for 20 minutes. The resulting precipitate was filtered offand washed with methanol (10 mL). The filtrate was concentrated to givethe title compound (13.2 g, 65%) as a solid. −ESI (M−H) 200.0; ¹H NMR(400 MHz, DMSO-d₆, δ): 8.43-8.45 (m, 1H), 8.05 (dd, J=8.8, 1.6 Hz, 1H),7.85 (dd, J=8.9, 0.9 Hz, 1H), 3.88 (s, 3H).

Step 2. 3-carbamoyl-1H-indazole-5-carboxylic acid

A suspension of methyl 3-cyano-1H-indazole-5-carboxylate (50.0 g, 249mmol) in methanol (1 L) was cooled to 10° C. A solution of urea hydrogenperoxide (241 g, 2.49 mol) in sodium hydroxide (1 L of 2.5 N) and water(100 mL) was added dropwise, maintaining an internal temperature below25° C. When the addition was complete, the ice bath was removed and thereaction was allowed to stir at room temperature for 16 hours. A smallamount of unreacted starting material was observed by HPLC. The reactionwas cooled to 15° C. and additional urea hydrogen peroxide (50 g) wasadded portionwise. Vigorous bubbling was noted. The reaction was allowedto stir for another 2 hours. The crude reaction was filtered to removethe solids present and the filtrate was concentrated to remove themethanol. The remaining solution was cooled in an ice bath and 6 Nhydrochloric acid (420 mL) was added dropwise to adjust the pH to 4. Thesolution was stirred for 20 minutes and the resulting tan solid wascollected by filtration and dried to give 57.2 g of crude product. Tothe crude was added acetonitrile (700 mL) and dichloromethane (700 mL)and the mixture was stirred at room temperature for 1 hour. The solidwas collected by filtration, washed with 1:1 acetonitrile:dichloromethane (400 mL) and dried to give the title compound (39.5 g,77%) as a tan solid. +ESI (M+H) 206.1; ¹H NMR (400 MHz, DMSO-d₆, δ):13.81 (s, 1H), 12.85 (br. s., 1H), 8.82 (d, J=0.8 Hz, 1H), 7.93 (dd,J=8.8, 1.6 Hz, 1H), 7.79-7.85 (m, 1H), 7.64 (d, J=8.6 Hz, 1H), 7.44 (s,1H).

Intermediate 10: 3-cyano-1H-indazole-5-carboxylic acid, shown below, wasprepared as follows

Methyl 3-cyano-1H-indazole-5-carboxylate (500 mg, 2.5 mmol) wasdissolved in methanol (12 mL) and 2 N aqueous lithium hydroxide (3.7 mL,7 mmol) was added. The reaction was stirred at room temperatureovernight. The reaction mixture was concentrated to remove the methanoland the residue was acidified to pH=4 with 1 N aqueous hydrochloricacid. The resulting yellow precipitate was collected by filtration,washed with water, and dried in a vacuum oven to provide the titlecompound (445 mg, 96%). −ESI (M−H) 186.4; ¹H NMR (400 MHz, DMSO-d₆, δ):13.17 (br. s., 1H), 8.42 (s, 1H), 8.05 (dd, J=8.8, 1.6 Hz, 1H), 7.83 (d,1H).

Intermediate 11: 3-cyano-1H-indazole-6-carboxylic acid, shown below, wasprepared as follows

Step 1: methyl 1H-indazole-6-carboxylate

To a solution of 1H-indazole-6-carboxylic acid (3.00 g, 18.5 mmol) inN,N-dimethylformamide (46 mL) was added sodium carbonate (2.06 g, 19.4mmol), followed by iodomethane (2.75 g, 1.21 mL, 19.4 mmol) dropwise.The mixture was stirred at room temperature overnight. The mixture waspoured into half saturated sodium bicarbonate and extracted with ethylacetate three times. The combined organic layers were washed with brine,dried over sodium sulfate, filtered and concentrated in vacuo to afforda brown oil. This residue was purified by flash column chromatography(12-100% ethyl acetate/heptanes) to afford methyl1H-indazole-6-carboxylate as a yellow solid (2.95 g, 90%). ¹H NMR (400MHz, CDCl₃, δ): 10.40 (br. s., 1H), 8.26 (s, 1H), 8.13 (s, 1H), 7.84 (d,J=8.4 Hz, 1H), 7.79 (d, J=8.4 Hz, 1H), 3.96 (s, 3H).

Step 2: methyl 3-iodo-1H-indazole-6-carboxylate

To a solution of methyl 1H-indazole-6-carboxylate (865 mg, 4.91 mmol) inN,N-dimethylformamide (12 mL) was added potassium hydroxide (840 mg,3.05 mmol) followed by iodine (1.54 g, 5.9 mmol). The mixture wasstirred at room temperature for 3 hours. Sodium bisulfate (30 mL of 5%aqueous) was added and the mixture was extracted with ethyl acetatetwice. The combined organic layers were washed with brine, dried oversodium sulfate, filtered and concentrated in vacuo. The residue waspurified via flash column chromatography (5-65% ethyl acetate/heptanes)to afford methyl 3-iodo-1H-indazole-6-carboxylate as a colorless solid(1.16 g, 78%). ¹H NMR (400 MHz, DMSO-d₆, δ): 13.84 (s, 1H), 8.13 (s,1H), 7.72 (d, J=8.4 Hz, 1H), 7.54 (d, J=8.6 Hz, 1H), 3.87 (s, 3H).

Step 3: methyl 3-cyano-1H-indazole-6-carboxylate

A mixture of methyl 3-iodo-1H-indazole-6-carboxylate (3.0 g, 9.9 mmol),zinc dust (400 mg, 6.11 mmol), zinc cyanide (2.0 g, 17.0 mmol),[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II), complexwith dichloromethane (1.15 g, 1.41 mmol), and copper (I) iodide (1.90 g,9.97 mmol) in dimethylacetamide (55 mL) was purged with nitrogen for 15minutes. The mixture was stirred at 120° C. for 15 hours. The reactionmixture was cooled, diluted with ethyl acetate (250 mL), and filteredthrough Celite, rinsing with ethyl acetate (100 mL). To the filtrate wasadded ˜400 mL of a solution of saturated aqueous ammonium chloride andconcentrated ammonium hydroxide (prepared by adding ammonium hydroxideto a saturated aqueous solution of ammonium chloride until pH=8). Themixture was stirred for 1 hour. The layers were then separated. Theorganic layer was washed with water and brine, dried over sodiumsulfate, filtered and concentrated in vacuo. To the residue was addedmethanol (40 mL) and the mixture was stirred overnight. The mixture wasfiltered and the solid was dried in vacuo to give methyl3-cyano-1H-indazole-6-carboxylate as a tan solid (1.47 g, 73%). ¹H NMR(400 MHz, DMSO-d₆, δ): 13.40 (br. s., 1H), 8.25 (s, 1H), 7.94 (d, J=8.6Hz, 1H), 7.83 (d, J=8.4 Hz, 1H), 3.88 (s, 3H).

Step 4: 3-cyano-1H-indazole-6-carboxylic acid

To a solution of methyl 3-cyano-1H-indazole-6-carboxylate (1.47 g, 7.31mmol) in methanol (36 mL) and tetrahydrofuran (20 mL) was added 2 Naqueous lithium hydroxide (16 mL, 32 mmol). The reaction was heated to50° C. for 72 hours. The reaction was cooled to room temperature andconcentrated. The residue was diluted with water and the pH was adjustedto 4 with 1 N aqueous hydrochloric acid. The resulting precipitate wasfiltered off, rinsed with water, and dried under vacuum to provide thetitle compound (500 mg, 37%) as a tan solid. +ESI (M+H) 188.2.

Intermediate 12: 1-methoxyisoquinoline-7-carboxylic acid, shown below,was prepared as follows

Step 1: 7-bromo-1-methoxyisoquinoline

7-Bromo-1-chloroisoquinoline (570 mg, 2.4 mmol) was combined withmethanol (10 mL) and sodium methoxide (25 wt % in methanol, 1.5 mL, 24mmol) in a microwave vial. The vial was sealed and heated to 130° C. for3 hours in a microwave. The reaction was concentrated. The crude residuewas taken up in ethyl acetate and washed with water and saturatedaqueous sodium bicarbonate. The aqueous layer was extracted two timeswith hot ethyl acetate. The combined organics were dried over sodiumsulfate, filtered, and concentrated to give the title compound (520 mg,93%). +ESI (M+H+1) 240.0; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.25-8.28 (m,1H), 8.04 (d, J=5.9 Hz, 1H), 7.86-7.89 (m, 2H), 7.40 (dd, J=6.0, 0.9 Hz,1H), 4.03 (s, 3H).

Step 2: methyl 1-methoxyisoquinoline-7-carboxylate

To a solution of 7-bromo-1-methoxyisoquinoline (520 mg, 2.2 mmol) inmethanol (30 mL) was added sodium acetate (517 mg, 6.30 mmol) and[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(11), complexwith dichloromethane (257 mg, 0.315 mmol). The mixture was evacuated andbackfilled with nitrogen three times. The reaction vessel was thenpressurized to 25 psi carbon monoxide. The reaction was heated to 70° C.and was agitated for 20 hours. The reaction was filtered, rinsing withmethanol. The filtrate was concentrated. The resulting residue was takenup in dichloromethane and the remaining solids were filtered off. Thefiltrate was concentrated and purified by flash column chromatography(0-100% ethyl acetate/heptanes) to give the title compound (443 mg, 93%)as a white solid. +ESI (M+H) 218.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.77(d, J=0.8 Hz, 1H), 8.20 (dd, J=8.6, 1.8 Hz, 1H), 8.13 (d, J=5.9 Hz, 1H),8.00 (d, J=8.6 Hz, 1H), 7.46 (d, J=5.9 Hz, 1H), 4.08 (s, 3H), 3.90 (s,3H).

Step 3: 1-methoxyisoquinoline-7-carboxylic acid

To a solution of methyl 1-methoxyisoquinoline-7-carboxylate (443 mg,2.04 mmol) in methanol (10 mL) was added 2 N aqueous lithium hydroxide(20 mL). The reaction was stirred at room temperature for 24 hours. Thereaction mixture was diluted with 1 N aqueous hydrochloric acid andethyl acetate. The layers were separated and the aqueous was extractedtwo more times with ethyl acetate. The combined organics were washedwith brine, dried over sodium sulfate, filtered, and concentrated toafford the title compound (414 mg, 100%) as a solid. +ESI (M+H) 204.1;¹H NMR (400 MHz, DMSO-d₆, δ): 13.24 (s, 1H), 8.76 (d, J=0.8 Hz, 1H),8.18 (dd, J=8.6, 1.8 Hz, 1H), 8.11 (d, J=5.9 Hz, 1H), 7.97 (d, J=8.4 Hz,1H), 7.45 (d, J=5.9 Hz, 1H), 4.07 (s, 3H).

Intermediate 13: 3-aminoisoquinoline-6-carboxylic acid, shown below, wasprepared as follows:

The title compound was prepared by a method analogous to that describedfor Intermediate 12, Steps 2-3, using 6-bromoisoquinolin-3-amine. +ESI(M+H) 189.0; ¹H NMR (400 MHz, DMSO-d₆, δ): 13.15 (br. s., 1H), 8.94 (s,1H), 8.20 (s, 1H), 7.91 (m, 1H), 7.62-7.59 (m, 1H), 6.78 (s, 1H), 6.14(s, 2H).

Intermediate 14: 3-amino-1H-indazole-5-carboxylic acid, shown below, wasprepared as follows

To a solution of 3-cyano-4-fluorobenzoic acid (980.0 mg, 5.94 mmol) inethanol (6 mL), was added hydrazine hydrate (0.89 mL, 17.8 mmol). Thereaction was heated at reflux for 3 hours. The reaction was cooled toroom temperature and ethanol was removed under reduced pressure. Theresultant yellow oil was taken up in water (50 mL) and basified with 1 Naqueous sodium hydroxide (5 mL). The solution was washed once with ethylacetate (25 mL). The aqueous phase was acidified to pH=3 with 6 Naqueous hydrochloric acid and was allowed to stir at room temperaturefor 1 hour. The resulting precipitate was collected by filtration anddried under vacuum to give the title compound (612 mg, 48%) as a pinksolid. +ESI (M+H) 178.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.42-8.47 (m,1H), 7.76 (dd, J=8.8, 1.6 Hz, 1H), 7.21 (d, J=8.8 Hz, 1H).

Intermediate 15: 3-amino-1H-indazole-6-carboxylic acid, shown below, wasprepared as follows

To a solution of 4-cyano-3-fluorobenzoic acid (500 mg, 3.0 mmol) inn-butanol (9 mL) was added hydrazine monohydrate (0.5 mL, 10 mmol). Thereaction was heated to 110° C. overnight. The reaction was cooled toroom temperature and the precipitate was collected by filtration. Thesolid was then dissolved in 1 N aqueous sodium hydroxide (2 mL) andextracted with ethyl acetate (2×). The aqueous layer was acidified topH=4 with 1 N aqueous hydrochloric acid. The resulting precipitate wascollected by filtration and dried under vacuum to provide the titlecompound (140 mg, 26%) as a red solid. +ESI (M+H) 178.2; ¹H NMR (400MHz, CD₃OD, δ): 7.99-8.01 (m, 1H), 7.73 (dd, J=8.4, 0.8 Hz, 1H), 7.61(dd, J=8.5, 1.3 Hz, 1H).

Intermediate 16: 2-methyl-3-oxo-2,3-dihydro-1H-indazole-5-carboxylicacid, shown below, was prepared as follows

Step 1: methyl 3-hydroxy-1H-indazole-5-carboxylate

3-hydroxy-1H-indazole-5-carboxylic acid (1.5 g, 8.4 mmol) was suspendedin methanol (17 mL). Concentrated hydrochloric acid (3.11 mL, 101 mmol)was added and the reaction was heated to 100° C. for 6 hours. Thereaction was concentrated to provide the title compound (1.60 g, 99%).+ESI (M+H) 193.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 12.00 (br. s., 1H), 8.35(s, 1H), 7.83 (dd, J=8.9, 1.7 Hz, 1H), 7.33 (dd, J=8.9, 0.7 Hz, 1H),3.82 (s, 3H).

Step 2: 1-ethyl 5-methyl 3-hydroxy-1H-indazole-1,5-dicarboxylate

Methyl 3-hydroxy-1H-indazole-5-carboxylate (1.60 g, 8.33 mmol) wassuspended in pyridine (10 mL). Ethyl chloroformate (0.90 mL, 9.3 mmol)was added slowly and the reaction was stirred at room temperature for 1hour. Additional ethyl chloroformate (0.30 mL, 3.1 mmol) was added andthe reaction was stirred for another 30 minutes. The reaction was pouredinto water (65 mL) and cooled in a refrigerator for 3 hours. The brownsolid was collected by filtration, rinsed with water, and dried undervacuum to give the title compound (1.75 g, 80%). +ESI (M+H) 265.1; ¹HNMR (400 MHz, CDCl₃, δ): 8.56 (s, 1H), 8.29 (d, J=7.8 Hz, 1H), 8.13 (br.s., 1H), 4.59 (q, J=7.0 Hz, 2H), 3.97 (s, 3H), 1.56 (t, J=7.0 Hz, 3H).

Step 3: 1-ethyl 5-methyl2-methyl-3-oxo-2,3-dihydro-1H-indazole-1,5-dicarboxylate

1-Ethyl 5-methyl 3-hydroxy-1H-indazole-1,5-dicarboxylate (1.75 g, 6.62mmol) was suspended in acetone (85 mL). Cesium carbonate (2.27 g, 6.95mmol) and methyl iodide (1.3 mL, 20 mmol) were added and the reactionwas stirred at reflux for 22 hours. The reaction was concentrated todryness and the residue was partitioned between dichloromethane (60 mL)and water (100 mL). The layers were separated and the aqueous wasextracted again with dichloromethane (60 mL). The combined organics weredried over magnesium sulfate, filtered, and concentrated. Purificationby flash column chromatography (7-60% ethyl acetate/heptanes) gave tworegioisomeric products.

1-ethyl 5-methyl 3-methoxy-1H-indazole-1,5-dicarboxylate (590 mg, 32%)as a white solid. ¹H NMR (400 MHz, CDCl₃, δ): 8.41 (dd, J=1.6, 0.8 Hz,1H), 8.22 (dd, J=9.2, 3.5 Hz, 1H), 8.14 (d, J=9.2 Hz, 1H), 4.57 (q,J=7.1 Hz, 2H), 4.20 (s, 3H), 3.95 (s, 3H), 1.51 (t, J=7.1 Hz, 3H).

1-ethyl 5-methyl2-methyl-3-oxo-2,3-dihydro-1H-indazole-1,5-dicarboxylate (699 mg, 38%)as a yellow solid. +ESI (M+H) 279.1; ¹H NMR (400 MHz, CDCl₃, δ): 8.56(dd, J=1.8, 0.6 Hz, 1H), 8.30 (dd, J=8.8, 1.8 Hz, 1H), 7.93 (d, J=8.8Hz, 1H), 4.50 (q, J=7.0 Hz, 2H), 3.94 (s, 3H), 3.67 (s, 3H), 1.48 (t,J=7.1 Hz, 3H).

Step 4: 2-methyl-3-oxo-2,3-dihydro-1H-indazole-5-carboxylic acid

1-Ethyl 5-methyl2-methyl-3-oxo-2,3-dihydro-1H-indazole-1,5-dicarboxylate (300 mg, 1.08mmol) was dissolved in ethanol (4 mL). Potassium hydroxide (485 mg, 8.62mmol) was added and the reaction was stirred at room temperature for 1.5hours. LCMS showed the reaction to be incomplete. An aqueous solution ofpotassium hydroxide (10 mL, 10 mmol, 1.0 M) was then added and thereaction was heated to 65° C. for 2 hours. The reaction was cooled toroom temperature and concentrated. The resulting orange solid wasdissolved in water and acidified with 1 N aqueous hydrochloric acid. Theprecipitate was collected by filtration and dried under vacuum to givethe title compound (158 mg, 76%) as a white solid. +ESI (M+H) 193.1; ¹HNMR (400 MHz, DMSO-d₆, δ): 12.75 (br. s., 1H), 11.06 (s, 1H), 8.15 (s,1H), 7.99 (dd, J=8.7, 1.5 Hz, 1H), 7.28 (d, J=8.6 Hz, 1H), 3.37 (s, 3H).

Intermediate 17: 3-methoxy-1H-indazole-5-carboxylic acid, shown below,was prepared as follows

The title compound was prepared by a method analogous to that describedfor Intermediate 16, using 1-ethyl 5-methyl3-methoxy-1H-indazole-1,5-dicarboxylate, the regioisomeric productformed in Step 3. +ESI (M+H) 193.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 12.65(br. s., 1H), 12.26 (s, 1H), 8.18 (s, 1H), 7.86 (dd, J=8.9, 1.5 Hz, 1H),7.38 (d, J=8.8 Hz, 1H), 3.99 (s, 3H).

Intermediate 18:7-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylic acid,shown below, was prepared as follows

Step 1: ethyl7-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylate

To a mixture of ethyl7-hydroxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylate(WO2009144554) (100 mg, 0.34 mmol) and potassium carbonate (95.1 mg,0.68 mmol) in N,N-dimethylformamide (1 mL) was added methyl iodide (32μL, 0.51 mmol). The reaction was stirred at room temperature overnight.The reaction was diluted with water and extracted with ethyl acetate(4×). The combined organic layers were washed with water and brine,dried over sodium sulfate, filtered, and concentrated to give the titlecompound (105 mg, 100%) as a yellow oil. +ESI (M+1-THP) 221.2; ¹H NMR(400 MHz, CDCl₃, δ): 8.07-8.10 (m, 2H), 7.43 (d, J=0.98 Hz, 1H), 6.24(dd, J=10.24, 2.44 Hz, 1H), 4.38 (q, J=7.15 Hz, 2H), 4.08 (dt, J=11.56,2.02 Hz, 1H), 4.04 (s, 3H), 3.70-3.78 (m, 1H), 2.54-2.66 (m, 1H),2.09-2.19 (m, 1H), 2.01-2.08 (m, 1H), 1.71-1.83 (m, 2H), 1.55-1.64 (m,1H), 1.41 (t, J=7.12 Hz, 3H).

Step 2: 7-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylicacid

To a solution of ethyl7-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylate (102mg, 0.33 mmol) in tetrahydrofuran (2 mL) was added 1 N aqueous lithiumhydroxide (0.67 mL, 0.67 mmol). The reaction was stirred at roomtemperature overnight. LCMS showed the reaction to be incomplete.Additional lithium hydroxide (0.35 mL, 2 M, 0.7 mmol) was added and thereaction was heated to 40° C. for 1 hour. The reaction was then leftstirring at room temperature for 70 hours. The tetrahydrofuran wasremoved in vacuo and the residue was acidified to pH=4 with 1 N aqueoushydrochloric acid. The solution was extracted with ethyl acetate (3×).The combined organics were washed with brine, dried over sodium sulfate,filtered, and concentrated to give the title compound (84 mg, 91%) as asolid. (M+1-THP) 193.2; ¹H NMR (400 MHz, CDCl₃, δ): 8.18 (d, J=1.37 Hz,1H), 8.12 (s, 1H), 7.46 (d, J=1.17 Hz, 1H), 6.26 (dd, J=10.15, 2.54 Hz,1H), 4.07-4.12 (m, 1H), 4.06 (s, 3H), 3.65-3.81 (m, 1H), 2.54-2.72 (m,1H), 2.10-2.22 (m, 1H), 2.01-2.10 (m, 1H), 1.71-1.85 (m, 2H), 1.57-1.67(m, 1H).

Intermediate 19: 2-methoxyquinoline-7-carboxylic acid, shown below, wasprepared as follows

Step 1: 7-(ethoxycarbonyl)quinoline 1-oxide

To a solution of ethyl quinoline-7-carboxylate (1.02 g, 5.05 mmol) indichloromethane (20 mL) was added peracetic acid (2.13 mL, 10.1 mmol, 32wt % in acetic acid). The reaction was stirred at room temperatureovernight. The reaction was partitioned between water anddichloromethane. The layers were separated and the aqueous was extractedwith dichloromethane (4×). The combined organics were washed with waterand brine, dried over sodium sulfate, filtered, and concentrated. Thesolid was concentrated from heptanes and ethyl acetate several times,then dried under vacuum to give the title compound (1.01 g, 92%) as ayellow solid. +ESI (M+H) 218.2; ¹H NMR (400 MHz, CDCl₃, δ): 9.40 (s,1H), 8.65 (d, J=6.05 Hz, 1H), 8.27 (dd, J=8.58, 1.56 Hz, 1H), 7.95 (d,J=8.39 Hz, 1H), 7.82 (d, J=8.58 Hz, 1H), 7.42 (dd, J=8.49, 6.15 Hz, 1H),4.47 (q, J=7.02 Hz, 2H), 1.45 (t, J=7.1 Hz, 3H).

Step 2: ethyl 2-methoxyquinoline-7-carboxylate

To a 0° C. solution of 7-(ethoxycarbonyl)quinoline 1-oxide (150 mg, 0.69mmol) and toluene-4-sulphonyl chloride (171 mg, 0.89 mmol) in methanol(5 mL) was added triethylamine (0.19 mL, 1.4 mmol).

The reaction was stirred at room temperature overnight. LCMS showed thereaction was incomplete. Additional triethylamine (0.05 mL) was addedand the reaction was stirred for another 4 hours. The reaction wasconcentrated and the residue partitioned between ethyl acetate andsaturated aqueous sodium carbonate. The layers were separated and theaqueous was extracted two more times with ethyl acetate. The combinedorganics were washed with brine, dried over sodium sulfate, filtered,and concentrated. Purification by flash column chromatography (0˜40%ethyl acetate/heptanes) gave the title compound (130 mg, 81%) as a paleyellow solid. +ESI (M+H) 232.2; ¹H NMR (400 MHz, CDCl₃, δ): 8.49-8.60(m, 1H), 7.95-8.05 (m, 2H), 7.75 (d, J=8.19 Hz, 1H), 6.98 (d, J=8.78 Hz,1H), 4.43 (q, J=7.22 Hz, 2H), 4.08 (s, 3H), 1.43 (t, J=7.12 Hz, 3H).

Step 3: 2-methoxyquinoline-7-carboxylic acid

To a solution of ethyl 2-methoxyquinoline-7-carboxylate (125 mg, 0.54mmol) in tetrahydrofuran (1.5 mL) was added 2 N aqueous lithiumhydroxide (0.81 mL, 1.6 mmol). The reaction was stirred at roomtemperature for 65 hours. The tetrahydrofuran was removed in vacuo andthe residue was acidified to pH=4 with 1 N aqueous hydrochloric acid.The mixture was diluted with water and the resulting precipitate wascollected by filtration and dried under vacuum to give the titlecompound (106 mg, 96%) as a white solid. +ESI (M+H) 204.2; ¹H NMR (400MHz, CDCl₃, δ): 8.64 (d, J=1.37 Hz, 1H), 8.01-8.04 (m, 1H), 8.01 (s,1H), 7.79 (d, J=8.58 Hz, 1H), 7.01 (d, J=8.78 Hz, 1H), 4.09 (s, 3H).

Intermediate 20: 2-(methylamino)quinoline-6-carboxylic acid, shownbelow, was prepared as follows

Step 1: ethyl quinoline-6-carboxylate

To a solution of quinoline-6-carboxylic acid (2.8 g, 16 mmol) in ethanol(100 mL) was added concentrated sulfuric acid (2 mL). The reaction washeated to reflux overnight. The solvent was evaporated to give a brownresidue that was taken up in ethyl acetate (150 mL). The mixture waswashed with water (2×30 mL), saturated aqueous sodium bicarbonate (2×30mL), and brine (2×30 mL). The organic layer was dried over sodiumsulphate, filtered, and concentrated to an oil. Purification by flashcolumn chromatography gave the title compound (2.0 g, 81%) as a brownsolid.

Step 2: 6-(ethoxycarbonyl)quinoline 1-oxide

To ethyl quinoline-6-carboxylate (3.2 g, 16 mmol) in dichloromethane(120 mL) was added meta-chloroperoxybenzoic acid (4.9 g, 0.024 mol)portionwise. The reaction was stirred at room temperature for 4 hours.The reaction was diluted with dichloromethane and washed with saturatedaqueous sodium carbonate (3×30 mL) and brine (2×40 mL). The organiclayer was dried over sodium sulfate, filtered, and concentrated.Purification by flash column chromatography gave the title compound(2.45 g, 71%) as a brown solid. ¹H NMR (400 MHz, CDCl₃, δ): 8.81-8.79(d, 1H), 8.62 (s, 2H), 8.35-8.33 (d, 1H), 7.87-7.85 (d, 1H), 7.39 (s,1H), 4.49-4.44 (q, 2H), 1.47-1.43 (t, 3H).

Step 3: ethyl 2-(methylamino)quinoline-6-carboxylate

Trifluoromethanesulfonic anhydride (1.92 mL, 11.4 mmol) was addeddropwise to a −70° C. solution of 6-(ethoxycarbonyl)quinoline 1-oxide(2.25 g, 10.4 mmol) in dichloromethane (150 mL). The mixture was stirredat −70° C. for 5 minutes. Then a solution of methylamine intetrahydrofuran (31 mL, 62 mmol, 2 M) was added dropwise. The mixturewas stirred at −70° C. for 5 minutes. The reaction was quenched withwater (20 mL). The layers were separated and the aqueous was extractedwith dichloromethane (3×30 mL). The combined organics were washed withbrine, dried over sodium sulfate, filtered, and concentrated.Purification by flash column chromatography gave the title compound (850mg, 35%) as a yellow solid. ¹H NMR (400 MHz, CDCl₃, δ): 8.33 (d, 1H),8.16-8.13 (m, 1H), 7.90-7.87 (d, 1H), 7.70-7.67 (d, 1H), 6.68 (d, 1H),5.30 (br. s., 1H), 4.43-4.38 (q, 2H), 3.13-3.12 (d, 3H), 1.44-1.40 (m,3H).

Step 4: 2-(methylamino)quinoline-6-carboxylic acid

Aqueous sodium hydroxide (4 mL, 8 mmol, 2 N) was added to a solution ofethyl 2-(methylamino)quinoline-6-carboxylate (850 mg, 3.7 mmol) inethanol (10 mL). The reaction was heated to 50° C. overnight. Ethanolwas removed in vacuo and the residue was acidified to pH=5 with 1 Naqueous hydrochloric acid. The resulting precipitate was collected byfiltration and dried under vacuum to give the title compound (710 mg,96%) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆, δ): 8.26 (d, 1H),7.96-7.93 (m, 2H), 7.50 (d, 1H), 7.43 (d, 1H), 6.81 (d, 1H), 2.91 (d,3H).

Intermediate 21: 7-(trifluoromethyl)-1H-indazole-5-carboxylic acid,shown below, was prepared as follows:

Step 1: 4-bromo-2-methyl-6-(trifluoromethyl)aniline

To a room temperature solution of 2-methyl-6-(trifluoromethyl)aniline(3.0 g, 17 mmol) in acetonitrile (85 mL) was added N-bromosuccinimide(3.0 g, 17 mmol) in small portions over 30 minutes. The reaction wasallowed to stir for 1 hour. The reaction was poured into a water/brinemixture and was extracted with ethyl acetate (3×). The combined organicswere dried over magnesium sulfate, filtered, and concentrated.Purification by flash column chromatography (0-40% ethylacetate/heptanes) gave the title compound (4.13 g, 95%) as a brown oil.¹H NMR (400 MHz, CDCl₃, δ): 7.42 (d, J=2.34 Hz, 1H), 7.31 (s, 1H), 2.17(s, 3H).

Step 2: 5-bromo-7-(trifluoromethyl)-1H-indazole

To a solution of 4-bromo-2-methyl-6-(trifluoromethyl)aniline (3.3 g, 13mmol) in toluene (65 mL) and glacial acetic acid (11.2 mL, 195 mmol) wasadded potassium acetate (10.2 g, 104 mmol) portionwise. After 15 minutesa large amount of precipitate had formed, hindering stirring of thereaction. The reaction was diluted with acetic acid (10 mL). Isoamylnitrite (1.92 mL, 14.3 mmol) was then added dropwise and the reactionwas stirred at room temperature for 3 hours. Additional isoamyl nitrite(0.5 mL, 3.7 mmol) was added and the reaction was left stirring for 15hours. The reaction was diluted with water (100 mL) and stirred for 1.5hours. The solution was partitioned between ethyl acetate and saturatedaqueous sodium bicarbonate. The layers were separated and the organicswere washed with brine, dried over magnesium sulfate, filtered, andconcentrated. Purification by flash column chromatography (5-50% ethylacetate/heptanes) gave the title compound (1.78 g, 52%) as a yellowpowder. −ESI (M−H+1) 264.9; ¹H NMR (400 MHz, CDCl₃, δ): 8.13 (s, 1H),8.09-8.11 (m, 1H), 7.76 (dd, J=1.66, 0.88 Hz, 1H).

Step 3: methyl 7-(trifluoromethyl)-1H-indazole-5-carboxylate

To a sealed tube was added[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II), complexwith dichloromethane (45.7 mg, 0.056 mmol),5-bromo-7-(trifluoromethyl)-1H-indazole (100 mg, 0.38 mmol),triethylamine (105 μL, 0.752 mmol), and methanol (2 mL). The tube wascapped and carbon monoxide was bubbled through for 5 minutes. Thereaction was then heated to 70° C. for 5 hours. The reaction was cooledto room temperature and left stirring for 2 days. The reaction wasconcentrated and purified by flash column chromatography (0-50% ethylacetate/heptanes) to give the title compound (64 mg, 69%) as a whitepowder. −ESI (M−H) 243.1; ¹H NMR (400 MHz, CDCl₃, δ): 8.72 (s, 1H), 8.37(s, 1H), 8.28 (s, 1H), 3.98 (s, 3H).

Step 4: 7-(trifluoromethyl)-1H-indazole-5-carboxylic acid

To a solution of methyl 7-(trifluoromethyl)-1H-indazole-5-carboxylate(62 mg, 0.25 mmol) in methanol (2 mL) and tetrahydrofuran (2 mL) wasadded 1 N aqueous lithium hydroxide (0.76 mL, 0.76 mmol). The reactionwas heated to 60° C. for 17 hours. The reaction was concentrated and theresidue was diluted with water and acidified to pH=3 with 1 N aqueoushydrochloric acid. The solution was extracted with dichloromethane (3×).The combined organics were washed with brine, dried over magnesiumsulfate, filtered, and concentrated to give the title compound (17 mg,29%) as an off-white powder. +ESI (M+H) 231.1.

Intermediate 22: 3-(methylamino)isoquinoline-6-carboxylic acid, shownbelow, was prepared as follows

Step 1: 6-bromo-N-methylisoquinolin-3-amine

To a solution of 6-bromoisoquinolin-3-amine (50.0 mg, 2.6 mmol) inN,N-dimethylformamide (10 mL) was added N,N-dimethylformamidedimethylacetal (2 mL). The reaction vessel was sealed and heated in aBiotage Smith Synthesizer microwave to 110° C. for 20 minutes. Sodiumtriacetoxyborohydride (59 mg, 0.28 mmol) was then added to the reactionmixture. The vial was resealed and heated again to 110° C. on a BiotageSmith Synthesizer microwave for 10 minutes. The reaction wasconcentrated. The residue was dissolved in ethyl acetate (50 mL) andwashed with brine (2×20 mL). The organics were dried over sodiumsulfate, filtered, and concentrated. Purification by flash columnchromatography gave the title compound (23 mg, 43%) as a white solid. ¹HNMR (400 MHz, CDCl₃, δ): 8.76 (s, 1H), 7.74 (s, 1H), 7.61 (d, 1H), 7.28(d, 1H), 6.40 (s, 1H), 5.09-5.07 (m, 1H), 2.97 (s, 3H).

Step 2: 3-(methylamino)isoquinoline-6-carboxylic acid

Methyl 3-(methylamino)isoquinoline-6-carboxylate was prepared by amethod analogous to that described in Step 3 of Intermediate 21, using6-bromo-N-methylisoquinolin-3-amine. To the crude material (580 mg, 2.7mmol) was added water (5 mL), methanol (5 mL), and lithium hydroxidemonohydrate (300 mg, 7 mmol). The mixture was stirred at roomtemperature overnight. The reaction was concentrated and the residue wasacidified to pH=5 with 1 N aqueous hydrochloric acid. The resultingresidue was dried under vacuum and purified by reversed-phase HPLC togive the title compound (512 mg, 89%) as a white solid. ¹H NMR (400 MHz,CD₃OD, δ): 8.81 (s, 1H), 8.23 (s, 1H), 7.80 (d, 1H), 7.72 (d, 1H), 6.70(s, 1H), 2.93 (s, 3H).

Intermediate 23: 2-(methylamino)quinoline-7-carboxylic acid, shownbelow, was prepared as follows

The title compound was prepared by a method analogous to that describedin Steps 3-4 of Intermediate 20, using 7-(ethoxycarbonyl)quinoline1-oxide. ¹H NMR (400 MHz, DMSO-d₆, δ): 8.08 (s, 1H), 7.90 (d, 1H),7.71-7.62 (m, 2H), 7.21 (s, 1H), 6.84 (d, 1H), 2.91 (d, 3H).

Intermediate 24: 5-methoxyquinoline-3-carboxylic acid, shown below, wasprepared as follows

Methyl 5-methoxyquinoline-3-carboxylate (Organic and BiomolecularChemistry, 7(12), 2612-2618; 2009) was saponified using aqueous lithiumhydroxide. +ESI (M+H) 203.9; ¹H NMR (400 MHz, DMSO-d₆, δ): 9.30 (d, 1H),9.03 (d, 1H), 7.84-7.80 (m, 1H), 7.66 (d, 1H), 7.15 (d, 1H), 4.04 (s,3H).

Intermediate 25: 2-(methylamino)-1H-benzo[d]imidazole-5-carboxylic acid,shown below, was prepared as follows

Step 1. methyl 2-(methylamino)-1H-benzo[d]imidazole-5-carboxylate

A mixture of 3,4-diaminobenzoic acid (15 g, 0.09 mol) andisothiocyanatomethane (6.6 g, 0.09 mol) was dissolved in tetrahydrofuran(90 mL). The reaction was heated at reflux for 3 hours and was thenconcentrated. The residue was poured into ice water. The resultingprecipitate was filtered, washed with water, and dried under vacuum togive methyl 4-amino-3-(3-methylthioureido)benzoate (12.0 g, 56%).

To the solid (12 g, 0.05 mol) was added ethanol (200 mL), followed bymethyl iodide (35.5 g, 0.25 mol). The reaction was heated to reflux andstirred overnight. The reaction was concentrated and the residue wasbasified with ammonium hydroxide. The solids were collected byfiltration and washed with water. Purification by column chromatography(9-25% ethyl acetate/petroleum ether) gave the title compound (2.9 g,28%) as a yellow solid. ¹H NMR (400 MHz, CDCl₃, δ): 8.37 (s, 1H),7.92-7.96 (m, 1H), 7.51 (d, J=8.4 Hz, 1H), 3.93 (s, 3H), 2.81 (s, 3H).

Step 2. 2-(methylamino)-1H-benzo[d]imidazole-5-carboxylic acid

3 N Aqueous hydrochloric acid (14 mL, 42 mmol) was added to methyl2-(methylamino)-1H-benzo[d]imidazole-5-carboxylate (2.9 g, 14 mmol) andthe reaction was stirred at reflux overnight. The reaction wasconcentrated to give the title compound (2.4 g, 90%) as a yellow solid.¹H NMR (400 MHz, CD₃OD, δ): 7.96-8.00 (m, 2H), 7.40 (d, J=8.4 Hz, 1H),3.10 (s, 3H).

Intermediate 26: 2-amino-1H-benzo[d]imidazole-5-carboxylic acid, shownbelow, was prepared as follows

A solution of cyanogen bromide (5.0 mL, 5 M in acetonitrile, 25 mmol)was added to a mixture of methyl 3,4-diaminobenzoate (3.0 g, 18 mmol) inwater (50 mL). The reaction was stirred at room temperature overnight.Aqueous ammonia (20 mL) and ethyl acetate (100 mL) were added to thereaction mixture and the layers were separated. The organics were driedover sodium sulfate, filtered, and concentrated. To the crude residuewas added 2 N aqueous hydrochloric acid (18 mL, 36.0 mmol) and themixture was heated at reflux overnight. The reaction was concentrated togive the title compound (2.90 g, 97%). ¹H NMR (400 MHz, DMSO-d₆, δ):8.75 (s, 2H), 7.84 (s, 1H), 7.77 (dd, J=1.2, 8.4 Hz, 1H), 7.38 (d, J=8.4Hz, 1H).

Intermediate 27: 1-(4-methoxybenzylamino)isoquinoline-7-carboxylic acid,shown below, was prepared as follows

Step 1: 1-oxo-1,2-dihydroisoquinoline-7-carboxylic acid

To a suspension of 7-bromoisoquinolin-1 (2H)-one (70 g, 0.31 mol) inN,N-dimethylformamide (1 L) was added copper cyanide (56 g, 0.63 mol).The reaction was heated to 180° C. for 2 hours. The reaction was cooledto room temperature and was diluted with water (1 L). The solution wasextracted with ethyl acetate (3×). The organics were dried over sodiumsulfate, filtered, and concentrated to give crude1-oxo-1,2-dihydroisoquinoline-7-carbonitrile (37 g). This crude materialwas taken up in ethanol (500 mL) and 1 N aqueous sodium hydroxide (400mL) was added. The mixture was heated to reflux and stirred for 2 hours.The reaction was cooled to room temperature and the pH was adjusted to˜2 with 1 N aqueous hydrochloric acid. The solids were collected byfiltration, rinsed with water, and dried under vacuum to give the titlecompound (35 g, 85%) as an off-white solid. ¹H NMR (400 MHz, DMSO-d₆,δ): 13.15 (br. s., 1H), 11.49 (s, 1H), 8.75 (s, 1H), 8.17-8.14 (m, 1H),7.75 (d, 1H), 7.34-7.29 (m, 1H), 6.62 (d, 1H).

Step 2: 1-chloroisoquinoline-7-carbonyl chloride

Phosphorous oxychloride (74 mL, 793 mmol) was added to1-oxo-1,2-dihydroisoquinoline-7-carboxylic acid (3.0 g, 20 mmol). Thereaction was heated to 90° C. for 5 hours. The reaction was concentratedto dryness. The material was taken up in dichloromethane (250 mL) andsaturated aqueous sodium bicarbonate (200 mL). The layers were separatedand the aqueous was extracted again with dichloromethane (100 mL). Thecombined organics were dried over magnesium sulfate, filtered, andconcentrated to give the title compound (3.0 g, 80%) as a yellow solid.+ESI (M+H) 227.1; ¹H NMR (400 MHz, CDCl₃, δ): 9.18-9.22 (m, 1H), 8.44(d, J=5.7 Hz, 1H), 8.32 (dd, J=8.8, 1.8 Hz, 1H), 7.95 (d, J=8.8 Hz, 1H),7.68 (d, J=5.7 Hz, 1H).

Step 3: ethyl 1-chloroisoquinoline-7-carboxylate

1-Chloroisoquinoline-7-carbonyl chloride (3.02 g, 13.4 mmol) wasdissolved in tetrahydrofuran (135 mL) and was cooled to 0° C. Ethanol(6.1 mL, 94 mmol) and triethylamine (2.05 mL, 14.7 mmol) were added. Thereaction was allowed to warm to room temperature and stir for 2 hours.The reaction mixture was partitioned between ethyl acetate (500 mL) andsaturated aqueous sodium bicarbonate (250 mL). The organic layer wasdried over magnesium sulfate, filtered, and concentrated to give thetitle compound (3.0 g, 96%) as a yellow solid. +ESI (M+H) 236.1; ¹H NMR(400 MHz, CDCl₃, δ): 9.06 (s, 1H), 8.30-8.39 (m, 2H), 7.89 (d, J=8.6 Hz,1H), 7.63 (d, J=5.7 Hz, 1H), 4.48 (q, J=7.1 Hz, 2H), 1.46 (t, J=7.1 Hz,3H).

Step 4: ethyl 1-(4-methoxybenzylamino)isoquinoline-7-carboxylate

To a solution of ethyl 1-chloroisoquinoline-7-carboxylate (548 mg, 2.32mmol) in N,N-dimethylformamide (9.3 mL) was added 4-methoxy-benzylamine(4.6 mL, 35 mmol) and potassium carbonate (5.14 g, 37.2 mmol). Thereaction was heated to 70° C. and stirred overnight. The reaction wascooled to room temperature and was diluted with ethyl acetate and water.The layers were separated and the aqueous was extracted twice with ethylacetate. The combined organics were washed with water and brine, driedover magnesium sulfate, filtered, and concentrated. Purification byflash column chromatography (0-35% ethyl acetate/heptanes) gave thetitle compound (430 mg, 55%) as a greenish oil. ¹H NMR (400 MHz, CDCl₃,δ): 8.49 (s, 1H), 8.16 (dd, J=8.6, 1.6 Hz, 1H), 8.09 (d, J=5.9 Hz, 1H),7.69 (d, J=8.6 Hz, 1H), 7.33-7.40 (m, 2H), 6.96 (d, J=5.9 Hz, 1H),6.87-6.93 (m, 2H), 5.67 (br. s., 1H), 4.76 (d, J=5.1 Hz, 2H), 4.41 (q, J=7.2 Hz, 2H), 3.81 (s, 3H), 1.37-1.43 (m, 3H).

Step 5: 1-(4-methoxybenzylamino)isoquinoline-7-carboxylic acid

To a solution of ethyl1-(4-methoxybenzylamino)isoquinoline-7-carboxylate (430 mg, 1.28 mmol)in methanol (8.5 mL) was added 6 N aqueous sodium hydroxide (1.1 mL, 6.4mmol). The reaction was stirred at room temperature overnight. Thereaction was concentrated. The residue was taken up in water andacidified with 1 N aqueous hydrochloric acid until a precipitate formed.The solid was collected by filtration and dried under vacuum to give thetitle compound (328 mg, 83%) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆, δ): 8.92 (s, 1H), 8.30 (t, J=5.8 Hz, 1H), 8.06 (dd, J=8.4,1.4Hz, 1H), 7.88 (d, J=5.7 Hz, 1H), 7.69 (d, J=8.6 Hz, 1H), 7.24-7.31(m, 2H), 6.88 (d, J=5.7 Hz, 1H), 6.79-6.85 (m, 2H), 4.62 (d, J=5.9 Hz,2H), 3.67 (s, 3H).

Intermediate 28: 3-methoxy-1H-indazole-6-carboxylic acid, shown below,was prepared as follows

Step 1: methyl 3-hydroxy-1H-indazole-6-carboxylate

3-Oxo-2,3-dihydro-1H-indazole-6-carboxylic acid (1.5 g, 8.4 mmol) wassuspended in methanol (17 mL). Concentrated hydrochloric acid (3.1 mL,101 mmol) was added and the reaction was heated to reflux for 24 hours.The reaction was concentrated to give the title compound (1.6 g, 100%).+ESI (M+H) 193.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 11.98 (br. s., 1H), 7.89(s, 1H), 7.72 (d, J=8.6 Hz, 1H), 7.50 (dd, J=8.5, 1.3 Hz, 1H), 3.85 (s,3H).

Step 2: 1-ethyl 6-methyl 3-hydroxy-1H-indazole-1,6-dicarboxylate

Methyl 3-hydroxy-1H-indazole-6-carboxylate (1.6 g, 8.3 mmol) wassuspended in pyridine (10 mL). Ethyl chloroformate (1.0 mL, 10 mmol) wasadded slowly and the reaction was allowed to stir at room temperaturefor 2 hours. The reaction was poured into water (65 mL) and cooled in arefrigerator for 4 hours. The resulting brown precipitate was collectedby filtration, rinsed with water, and dried under vacuum to give thetitle compound (1.35 g, 61%) as a beige solid. +ESI (M+H) 265.0; ¹H NMR(400 MHz, CDCl₃, δ): 8.80 (d, J=6.0 Hz, 1H), 8.01 (dd, J=8.2, 1.2 Hz,1H), 7.88 (d, J=8.6 Hz, 1H), 4.60 (q, J=7.0 Hz, 2H), 3.98 (s, 3H), 1.57(t, J=7.1 Hz, 3H).

Step 3: 1-ethyl 6-methyl 3-methoxy-1H-indazole-1,6-dicarboxylate

1-Ethyl 6-methyl 3-hydroxy-1H-indazole-1,6-dicarboxylate (1.35 g, 5.11mmol) was suspended in acetone (65 mL). Cesium carbonate (1.75 g, 5.36mmol) and methyl iodide (1.0 mL, 15 mmol) were added and the reactionwas heated to reflux for 23 hours. The reaction was concentrated todryness. The residue was taken up in dichloromethane (100 mL) and water(100 mL). The layers were separated and the aqueous was extracted againwith dichloromethane. The combined organics were dried over magnesiumsulfate, filtered, and concentrated. Purification by flash columnchromatography gave two regioisomeric products.

1-ethyl 6-methyl 3-methoxy-1H-indazole-1,6-dicarboxylate (444 mg, 31%)as a white solid. ¹H NMR (400 MHz, CDCl₃, δ): 8.78 (s, 1H), 7.96 (dd,J=8.2, 1.4 Hz, 1H), 7.70 (dd, J=8.2, 0.8 Hz, 1H), 4.57 (q, J=7.2 Hz,2H), 4.19 (s, 3H), 3.96 (s, 3H), 1.51 (t, J=7.1 Hz, 3H).

1-ethyl 6-methyl2-methyl-3-oxo-2,3-dihydro-1H-indazole-1,6-dicarboxylate (514 mg, 36%)as a yellow solid. ¹H NMR (400 MHz, CDCl₃, δ): 8.56 (s, 1H), 8.00 (m,1H), 7.92 (d, J=8.6 Hz, 1H), 4.49 (q, J=7.2 Hz, 2H), 3.97 (s, 3H), 3.69(s, 3H), 1.49 (t, J=7.1 Hz, 3H).

Step 4: 3-methoxy-1H-indazole-6-carboxylic acid

1-Ethyl 6-methyl 3-methoxy-1H-indazole-1,6-dicarboxylate (444 mg, 1.60mmol) was suspended in ethanol (5 mL). An aqueous solution of potassiumhydroxide (16 mL, 16 mmol, 1 M) was added and the reaction was heated to65° C. and stirred for 1.5 hours. The reaction was cooled to roomtemperature and concentrated. The residue was taken up in water and thesolution was acidified with 1 N aqueous hydrochloric acid until aprecipitate formed. The solid was collected by filtration, rinsed withwater, and dried under vacuum to give the title compound (232 mg, 76%)as an orange solid. +ESI (M+H) 193.2; ¹H NMR (400 MHz, DMSO-d₆, δ):12.22 (s, 1H), 7.90-7.94 (m, 1H), 7.64 (d, J=8.4 Hz, 1H), 7.53 (dd,J=8.4, 1.4 Hz, 1H), 3.99 (s, 3H).

Intermediate 29: 3-(trifluoromethyl)-1H-indazole-5-carboxylic acid,shown below, was prepared as follows

The title compound was prepared by a method analogous to that describedin Steps 3-4 of Intermediate 21, using5-bromo-3-(trifluoromethyl)-1H-indazole. +ESI (M+H) 231.1.

Intermediate 30: 1-(4-methoxybenzylamino)isoquinoline-6-carboxylic acid,shown below, was prepared as follows

Step 1: 1-oxo-1,2-dihydroisoquinoline-6-carboxylic acid

A mixture of 6-bromoisoquinolin-1 (2H)-one (30 g, 0.134 mol),triethylamine (17.6 g, 0.174 mol), palladium(II) chloride (0.24 g, 1.34mmol) and 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (0.84 g, 1.34mmol) in methanol (300 mL) was pressurized with 2 MPa of carbonmonoxide. The reaction was heated to 100° C. and stirred for 12 hours.The reaction mixture was filtered through Celite and concentrated. Theresidue was washed with water and the solid was dried under vacuum togive crude methyl 1-oxo-1,2-dihydroisoquinoline-6-carboxylate (23.8 g,95.2%) as a yellow solid. The solid was diluted with tetrahydrofuran(200 mL) and water (200 mL). To this mixture was added lithium hydroxide(16.8 g, 0.4 mol) and the reaction was stirred at room temperature for 4hours. The reaction mixture was washed with ethyl acetate (3×) and thesewashings were discarded. The aqueous layer was acidified with 4 Naqueous hydrochloric acid to pH=5. The resulting precipitate wascollected by filtration and dried under vacuum to give the titlecompound (11.3 g, 49%) as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆, δ):11.48 (s, 1H), 8.24 (d, 2H), 7.93 (d, 1H), 7.22 (d, 1H), 6.68 (d, 1H).

Step 2: 1-(4-methoxybenzylamino)isoquinoline-6-carboxylic acid

The title compound was prepared by a method analogous to that describedin Steps 2-5 of Intermediate 27, using1-oxo-1,2-dihydroisoquinoline-6-carboxylic acid. +ESI (M+H) 309.2; ¹HNMR (400 MHz, CD₃OD, δ): 8.37 (d, J=1.56 Hz, 1H), 8.34 (d, J=8.78 Hz,1H), 8.12 (dd, J=8.68, 1.66 Hz, 1H), 7.67 (d, J=6.44 Hz, 1H), 7.29-7.36(m, 2H), 7.15 (d, J=6.24 Hz, 1H), 6.86-6.93 (m, 2H), 4.73 (s, 2H), 3.76(s, 3H).

Intermediate 31: 1-(methylamino)isoquinoline-7-carboxylic acid, shownbelow, was prepared as follows:

Step 1: ethyl 1-(methylamino)isoquinoline-7-carboxylate

A solution of methylamine in tetrahydrofuran (30 mL, 60 mmol, 2 M) wasadded to ethyl 1-chloroisoquinoline-7-carboxylate (formed in Step 3 ofIntermediate 27) (705 mg, 2.99 mmol) in a sealed tube. The reaction washeated to 60° C. and stirred overnight. LCMS indicated the reaction wasnot complete. Additional methylamine (10 mL, 20 mmol, 2 M in THF) wasadded and the reaction was heated to 60° C. for another 18 hours. Thereaction was cooled to room temperature and concentrated. The residuewas partitioned between water and dichloromethane. The organic layer wasdried over magnesium sulphate, filtered, and concentrated. Purificationby flash column chromatography (25-65% ethyl acetate/heptanes) gave thetitle compound (584 mg, 85%) as a yellow oil that solidified uponstanding. +ESI (M+H) 231.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.83-8.94 (m,1H), 8.07 (dd, J=8.58, 1.56 Hz, 1H), 7.99 (d, J=5.85 Hz, 1H), 7.89 (d,J=4.49 Hz, 1H), 7.77 (d, J=8.58 Hz, 1H), 6.92 (d, J=5.07 Hz, 1H), 4.38(q, J=7.02 Hz, 2H), 2.97 (d, J=4.49 Hz, 3H), 1.38 (t, J=7.12 Hz, 3H).

Step 2: 1-(methylamino)isoquinoline-7-carboxylic acid

The title compound was prepared by a method analogous to that describedin Step 3 of Intermediate 19, using ethyl1-(methylamino)isoquinoline-7-carboxylate. +ESI (M+H) 203.1; ¹H NMR (400MHz, DMSO-d₆, δ): 13.03 (br. s., 1H), 8.87 (s, 1H), 8.06 (dd, J=8.51,1.47 Hz, 1H), 7.97 (d, J=5.67 Hz, 1H), 7.85 (d, J=4.50 Hz, 1H), 7.75 (d,J=8.41 Hz, 1H), 6.91 (d, J=5.87 Hz, 1H), 2.95 (d, J=4.50 Hz, 3H).

Intermediate 32: 3-(trifluoromethyl)-1H-indazole-6-carboxylic acid,shown below, was prepared as follows

Step 1: 1-(4-bromo-2-fluorophenyl)-2,2,2-trifluoroethanol

To a 0° C. solution of 4-bromo-2-fluorobenzaldehyde (1.00 g, 4.93 mmol)in tetrahydrofuran (50 mL) was added trimethylsilyl trifluoromethane(0.77 mL, 4.9 mmol) dropwise over 5 minutes. The reaction was stirred at0° C. for 10 minutes. Then tetrabutylammonium fluoride (0.49 mL, 0.49mmol, 1 M in tetrahydrofuran) was slowly added and the reaction wasallowed to gradually warm to room temperature and stir for 3 days. Thereaction was concentrated and the residue was taken up indichloromethane. The solution was washed once with 1 N aqueoushydrochloric acid and once with brine. The organics were dried overmagnesium sulfate, filtered, and concentrated. Purification by columnchromatography (0-50% ethyl acetate/heptanes) gave the title compound(1.0 g, 75%) as a clear oil. ¹H NMR (400 MHz, CDCl₃, δ): 7.48 (d, J=7.61Hz, 1H), 7.39 (d, J=1.76 Hz, 1H), 7.29 (dd, J=9.56, 1.95 Hz, 1H),5.33-5.40 (m, 1H), 2.70 (d, J=5.46 Hz, 1H).

Step 2: 1-(4-bromo-2-fluorophenyl)-2,2,2-trifluoroethanone

To a solution of 1-(4-bromo-2-fluorophenyl)-2,2,2-trifluoroethanol (1.09g, 3.99 mmol) in ethyl acetate (30 mL) was added 2-iodoxybenzoic acid(2.28 g, 7.97 mmol). The reaction was heated to reflux overnight. Thereaction was cooled to room temperature and diluted with heptanes (30mL). The mixture was filtered through Celite and the filtrate wasconcentrated to give the title compound (1.03 g, 95%) as a pale yellowoil. ¹H NMR (400 MHz, CDCl₃, δ)_(:) 7.44 (dd, J=10.15, 1.56 Hz, 1H),7.48 (m, 1H), 7.76 (m, 1H).

Step 3: 6-bromo-3-(trifluoromethyl)-1H-indazole

Hydrazine hydrate (3.5 mL, 45 mmol) was added to a solution of1-(4-bromo-2-fluorophenyI)-2,2,2-trifluoroethanone (1.00 g, 3.69 mmol)in 1-butanol (15 mL). The reaction was heated to reflux for 5 hours,then cooled to room temperature and left stirring overnight. Thereaction was diluted with water (50 mL) and extracted with ethyl acetate(3×). The combined organics were washed with brine, dried over magnesiumsulfate, filtered, and concentrated. Purification by flash columnchromatography (0-50% ethyl acetate/heptanes) gave the title compound(310 mg, 32%) as an off-white solid. ¹H NMR (400 MHz, CDCl₃, δ): 7.42(dd, J=8.58, 1.56 Hz, 1H), 7.72 (d, J=8.58 Hz, 1H), 7.75 (dd, J=1.56,0.78 Hz, 1H).

Step 4: 3-(trifluoromethyl)-1H-indazole-6-carboxylic acid

The title compound was prepared by a method analogous to that describedin Steps 3-4 of Intermediate 21, using6-bromo-3-(trifluoromethyl)-1H-indazole. −ESI (M−H) 229.1.

Intermediate 33: 2-methyl-3-oxo-2,3-dihydro-1H-indazole-6-carboxylicacid, shown below, was prepared as follows

To a suspension of 1-ethyl 6-methyl2-methyl-3-oxo-2,3-dihydro-1H-indazole-1,6-dicarboxylate (formed in Step3 of Intermediate 28) (514 mg, 1.85 mmol) in ethanol (6 mL) was added 1N aqueous potassium hydroxide (18.5 mL, 18.5 mmol). The reaction washeated to 65° C. for 1.5 hours. The reaction was cooled to roomtemperature and concentrated to dryness. The residue was taken up inwater and acidified with 1 N aqueous hydrochloric acid until aprecipitate formed. The solid was collected by filtration and driedunder vacuum to give the title compound (196 mg, 55%) as a brown solid.¹H NMR (400 MHz, DMSO-d₆, δ): 13.12 (br. s., 1H), 10.61 (br. s., 1H),7.76 (s, 1H), 7.70 (d, J=8.2 Hz, 1H), 7.60 (dd, J=8.2, 1.2 Hz, 1H), 3.38(s, 3H).

Intermediate 34: 3-chloro-1H-pyrrolo[3,2-b]pyridine-6-carboxylic acid,shown below, was prepared as follows

Step 1: methyl 3-chloro-1H-pyrrolo[3,2-b]pyridine-6-carboxylate

To a 0° C. solution of methyl 1H-pyrrolo[3,2-b]pyridine-6-carboxylate(1.00 g, 5.68 mmol) in N,N-dimethylformamide (15 mL) was addedN-chlorosuccinimide (895 mg, 5.96 mmol). The reaction was allowed togradually warm to room temperature and stir overnight. The reaction wasdiluted with water (125 mL) and stirred for 20 minutes. The resultingsolid was collected by filtration, washed with water, and dried undervacuum to give the title compound (1.11 g, 93%) as an orange powder.+ESI (M+H) 211.0; ¹H NMR (400 MHz, DMSO-d₆, δ): 11.99 (br. s., 1H), 8.92(d, J=2.0 Hz, 1H), 8.31 (d, J=1.8 Hz, 1H), 8.08 (d, J=3.1 Hz, 1H), 3.88(s, 3H).

Step 2: 3-chloro-1H-pyrrolo[3,2-b]pyridine-6-carboxylic acid

Methyl 3-chloro-1H-pyrrolo[3,2-b]pyridine-6-carboxylate (1.10 g, 5.22mmol) was suspended in 1,4-dioxane (25 mL) and 6 N aqueous hydrochloricacid (8.7 mL) was added. The reaction was allowed to stir at roomtemperature overnight. The reaction was then concentrated to give thetitle compound (1.2 g, 100%). +ESI (M+H) 197.1; ¹H NMR (400 MHz,DMSO-d₆, δ): 12.50 (br. s., 1H), 8.92 (d, J=1.6 Hz, 1H), 8.46 (br. s.,1H), 8.19 (br. s., 1H).

Intermediate 35: 3-(methylamino)-1H-indazole-6-carboxylic acid, shownbelow, was prepared as follows

Step 1: 4-bromo-2-fluoro-N-methylbenzothioamide

A mixture of 4-bromo-2-fluoro-N-methylbenzamide (500 mg, 2 mmol) andLawesson reagent (872 mg, 2.16 mmol) in toluene (10 mL) was heated to100° C. and stirred for 4 hours. The reaction was cooled to roomtemperature, diluted with toluene, and filtered. The filtrate wasconcentrated and purification of the residue by flash columnchromatography (0-20% ethyl acetate/heptanes) gave the title compound(520 mg, 97%) as a yellow solid. +ESI (M+H+1) 250.1; ¹H NMR (400 MHz,CDCl₃, δ): 8.09 (t, J=8.58 Hz, 1H), 8.03 (br. s., 1H), 7.35 (dd, J=8.19,2.15 Hz, 1H), 7.27 (dd, J=11.41, 1.85 Hz, 1H), 3.36 (dd, J=4.88, 0.78Hz, 3H).

Step 2: 6-bromo-N-methyl-1H-indazol-3-amine

Anhydrous hydrazine (0.25 mL, 8.1 mmol) was added to a solution of4-bromo-2-fluoro-N-methylbenzothioamide (200 mg, 0.8 mmol) indimethylsulfoxide (2.5 mL). The reaction was heated to 100° C. andstirred for 2 hours. The reaction was cooled to room temperature anddiluted with ethyl acetate and water. The layers were separated and theaqueous was extracted with ethyl acetate (3×). The combined organicswere washed with saturated aqueous sodium carbonate and brine, driedover sodium sulfate, filtered, and concentrated. Purification by flashcolumn chromatography (20-100% ethyl acetate/heptanes) gave the titlecompound (98 mg, 54%) as a white solid. +ESI (M+H+1) 228.0; ¹H NMR (400MHz, CD₃OD, δ): 7.52 (d, J=8.58 Hz, 1H), 7.43 (s, 1H), 7.04 (d, J=8.39Hz, 1H), 2.94 (s, 3H).

Step 3: methyl 3-(methylamino)-1H-indazole-6-carboxylate

The title compound was prepared by a method analogous to that describedin Step 2 of Intermediate 12, using 6-bromo-N-methyl-1H-indazol-3-amine.+ESI (M+H) 206.2; ¹H NMR (400 MHz, CD₃OD, δ): 7.95 (t, J=1.17 Hz, 1H),7.67 (dd, J=8.39, 0.78 Hz, 1H), 7.55 (dd, J=8.49, 1.27 Hz, 1H), 3.90 (s,3H), 2.96 (s, 3H).

Step 4: 3-(methylamino)-1H-indazole-6-carboxylic acid

To a solution of methyl 3-(methylamino)-1H-indazole-6-carboxylate (30.0mg, 0.15 mmol) in 1,4-dioxane (0.2 mL) was added 3 N aqueoushydrochloric acid (0.2 mL, 0.6 mmol). The mixture was heated to 100° C.for 2 hours. The reaction was concentrated and dried under vacuum togive the title compound (33 mg, 99%) as a tan solid. +ESI (M+H) 192.1;¹H NMR (400 MHz, CD₃OD, δ): 8.09 (s, 1H), 7.98 (dd, J=8.58, 0.78 Hz,1H), 7.85 (dd, J=8.58, 1.37 Hz, 1H), 3.12 (s, 3H).

Intermediate 36: 3-methoxyisoquinoline-7-carboxylic acid, shown below,was prepared as follows:

Step 1: 7-bromo-3-methoxyisoquinoline

A mixture of 7-bromo-3-chloroisoquinoline (100 mg, 0.4 mmol) and sodiummethoxide (113 mg, 2.1 mmol) in diglyme (1 mL) was heated to 150° C. for1 hour. The reaction was cooled to room temperature and diluted withtoluene and water. The layers were separated and the aqueous layer wasextracted with toluene (3×). The combined organics were washed withwater and brine, dried over sodium sulfate, filtered, and concentratedto an oil. The oil was dried under vacuum overnight to give the titlecompound (83 mg, 85%) as a yellow solid. +ESI (M+H+1) 240.1; ¹H NMR (400MHz, CDCl₃, δ): 8.87 (s, 1H), 8.01-8.05 (m, 1H), 7.58-7.64 (m, 1H),7.53-7.58 (m, 1H), 6.97 (s, 1H), 4.02 (s, 3H).

Step 2: 3-methoxyisoquinoline-7-carboxylic acid

The title compound was prepared by a method analogous to that describedin Steps 3-4 of Intermediate 21, using 7-bromo-3-methoxyisoquinoline.+ESI (M+H) 204.2; ¹H NMR (400 MHz, CD₃OD, δ): 9.08 (s, 1H), 8.71 (s,1H), 8.14 (dd, J=8.78, 1.56 Hz, 1H), 7.83 (d, J=8.78 Hz, 1H), 7.17 (s,1H), 4.02 (s, 3H).

Intermediate 37: 1-(methylamino)isoquinoline-6-carboxylic acid, shownbelow, was prepared as follows

Step 1: ethyl 1-chloroisoquinoline-6-carboxylate

The title compound was prepared by a method analogous to that describedin Steps 1-3 of Intermediate 27, using 6-bromoisoquinolin-1 (2H)-one. ¹HNMR (400 MHz, CDCl₃, δ): 8.56 (d, J=1.6 Hz, 1 H), 8.38 (d, J=8.8 Hz,1H), 8.34 (d, J=5.7 Hz, 1H), 8.25 (dd, J=8.8, 1.6 Hz, 1H), 7.70 (d,J=6.0 Hz, 1H), 4.47 (q, J=7.0 Hz, 2H), 1.45 (t, J=7.1 Hz, 3H).

Step 2: ethyl 1-(methylamino)isoquinoline-6-carboxylate

The title compound was prepared by a method analogous to that describedin Step 1 of Intermediate 31, using ethyl1-chloroisoquinoline-6-carboxylate. +ESI (M+H) 231.1; ¹H NMR (400 MHz,CDCl₃, δ): 8.39 (s, 1H), 8.06-8.14 (m, 2H), 8.00 (d, J=5.9 Hz, 1H), 7.02(d, J=6.0 Hz, 1H), 4.44 (q, J=7.3 Hz, 2H), 3.25 (d, J=4.7 Hz, 3H), 1.43(t, J=7.1 Hz, 3H).

Step 3: 1-(methylamino)isoquinoline-6-carboxylic acid

To a suspension of ethyl 1-(methylamino)isoquinoline-6-carboxylate (150mg, 0.65 mmol) in ethanol (2.5 mL) was added 1 N aqueous potassiumhydroxide (6.5 mL, 6.5 mmol). The reaction was heated to 65° C. for 1.5hours. The reaction was cooled to room temperature and concentrated todryness. The solid was dissolved in water and the solution was acidifiedwith 1 N HCl. The mixture was concentrated. The solid was dissolved inwater (50 mL) and extracted twice with 2-butanol (50 mL). The combinedorganics were washed with brine (20 mL), dried over magnesium sulfate,filtered, and concentrated to give the title compound (95 mg, 72%) as awhite solid. +ESI (M+H) 203.2; ¹H NMR (400 MHz, DMSO-d₆, δ): 13.07(br.s., 1H), 10.25 (d, J=4.9 Hz, 1H), 8.74 (d, J=8.8 Hz, 1H), 8.51 (s,1H), 8.15 (dd, J=8.6, 1.8 Hz, 1H), 7.67 (d, J=6.8 Hz, 1H), 7.35 (d,J=7.0 Hz, 1H), 3.15 (d, J=4.7 Hz, 3H).

Intermediate 38: 1-methoxyisoquinoline-6-carboxylic acid

A solution of sodium methoxide was prepared by slowly adding sodiummetal (870 mg, 37 mmol) to methanol (25 mL) with stirring. After all ofthe sodium metal had reacted, this solution was added to ethyl1-chloroisoquinoline-6-carboxylate (440 mg, 1.9 mmol). The resultingsuspension was heated to reflux and stirred for 3 days. The reactionmixture was cooled to room temperature and concentrated. The residue waspartitioned between water and ethyl acetate. The layers were separatedand the aqueous layer was acidified with 1 N aqueous hydrochloric aciduntil a precipitate formed. The solid was collected by filtration anddried under vacuum to give the title compound (294 mg, 78%) as a whitesolid. ¹H NMR (400 MHz, DMSO-d₆, δ): 8.50 (d, J=1.2 Hz, 1H), 8.23 (d,J=8.8 Hz, 1H), 8.02-8.10 (m, 2H), 7.54 (d, J=6.0 Hz, 1H), 4.05 (s, 3H).

Intermediate 39: 3-(methylamino)-1H-indazole-5-carboxylic acid, shownbelow, was prepared as follows:

Step 1: 5-bromo-2-fluoro-N-methylbenzamide

To a mixture of 5-bromo-2-fluorobenzoic acid (200 mg, 0.91 mmol) indichloromethane (5 mL) was added oxalyl chloride (0.16 mL, 1.8 mmol),followed by 1 drop of N,N-dimethylformamide. The reaction was stirred atroom temperature for 1.5 hours. The reaction was concentrated and theresulting residue was dissolved in dichloromethane (3 mL) and cooled to0° C. Methylamine (2.3 mL, 5 mmol, 2 M in tetrahydrofuran) was added andthe reaction was allowed to stir at 0° C. for 30 minutes. The reactionwas quenched with water and the mixture was concentrated. The residuewas diluted with water and the resulting solids were filtered, rinsedwith water, and dried under vacuum to give the title compound (196.6 mg,93%) as a white solid. +ESI (M+H+1) 234.1; ¹H NMR (400 MHz, CDCl₃, δ);8.22 (dd, J=6.83, 2.73 Hz, 1H), 7.55 (ddd, J=8.68, 4.49, 2.63 Hz, 1H),7.00 (dd, J=11.32, 8.58 Hz, 1H), 6.67 (br. s., 1H), 3.02 (dd, J=4.88,1.17 Hz, 3H).

Step 2: 5-bromo-2-fluoro-N-methylbenzothioamide

A mixture of 5-bromo-2-fluoro-N-methylbenzamide (500 mg, 2 mmol) andLawesson's reagent (872 mg, 2.16 mmol) in toluene (10 mL) was heated to100° C. and stirred for 3.5 hours. The reaction was cooled to roomtemperature, diluted with toluene, and filtered. The filtrate wasconcentrated and purified by flash column chromatography (0-20% ethylacetate/heptanes) to give the title compound (494 mg, 92%) as a yellowoil that solidified upon standing. +ESI (M+H+1) 250.1; ¹H NMR (400 MHz,CDCl₃, δ): 8.20 (dd, J=6.93, 2.63 Hz, 1H), 8.06 (br.s., 1H), 7.47 (ddd,J=8.73, 4.44, 2.63 Hz, 1H), 6.95 (dd, J=11.12, 8.78 Hz, 1H), 3.32 (dd,J=4.88, 0.78 Hz, 3H).

Step 3: 5-bromo-N-methyl-1H-indazol-3-amine

A mixture of 5-bromo-2-fluoro-N-methylbenzothioamide (480 mg, 1.9 mmol)and anhydrous hydrazine (0.61 mL, 19 mmol) in dimethylsulfoxide (6 mL)was heated to 80° C. and stirred for 1 hour. The temperature wasincreased to 100° C. and the reaction was stirred for 40 minutes. Thetemperature was increased further to 130° C. and the reaction wasstirred for another 45 minutes. The reaction was cooled to roomtemperature and diluted with ethyl acetate and brine. The layers wereseparated and the aqueous was extracted with ethyl acetate (4×). Thecombined organics were washed with water and brine, dried over sodiumsulfate, filtered, and concentrated. Purification by flash columnchromatography (20-70% ethyl acetate/heptanes) gave the title compound(103 mg, 23%) as a white solid. +ESI (M+H+1) 228.0; ¹H NMR (400 MHz,CD₃OD, δ): 7.78 (dd, J=1.85, 0.68 Hz, 1H), 7.29-7.40 (m, 1H), 7.17 (dd,J=8.88, 0.68 Hz, 1H), 2.94 (s, 3H).

Step 4: methyl 3-(methylamino)-1H-indazole-5-carboxylate

The title compound was prepared by a method analogous to that describedin Step 3 of Intermediate 21, using 5-bromo-N-methyl-1H-indazol-3-amine.+ESI (M+H) 206.2; ¹H NMR (400 MHz, CD₃OD, δ): 8.44 (dd, J=1.56, 0.78 Hz,1H), 7.92 (dd, J=8.78, 1.56 Hz, 1H), 7.26 (dd, J=8.78, 0.78 Hz, 1H),3.88 (s, 3H), 2.96 (s, 3H).

Step 5: 3-(methylamino)-1H-indazole-5-carboxylic acid

Methyl 3-(methylamino)-1H-indazole-5-carboxylate (60.0 mg, 0.29 mmol)was dissolved in 1,4-dioxane (0.5 mL). 3N Aqueous hydrochloric acid (0.3mL, 0.9 mmol) was added and the reaction was heated to 100° C. for 11.5hours. The heat was removed and the reaction was left stirring at roomtemperature overnight. The reaction was concentrated to give the titlecompound (63 mg, 95%) as a tan solid. +ESI (M+H) 192.1; ¹H NMR (400 MHz,CD₃OD, δ): 8.61 (d, J=0.78 Hz, 1H), 8.19 (dd, J=8.80, 1.57 Hz, 1H), 7.38(d, J=8.80 Hz, 1H), 3.02 (s, 3H).

Intermediate 40: 3-aminoisoquinoline-7-carboxylic acid, shown below, wasprepared as follows

The title compound was prepared by a method analogous to that describedin Steps 3-4 of Intermediate 21, using 7-bromoisoquinolin-3-amine. +ESI(M+H) 189.2; ¹H NMR (400 MHz, CD₃OD, δ): 8.87 (s, 1H), 8.52 (d, J=0.78Hz, 1H), 7.98 (dd, J=8.78, 1.76 Hz, 1H), 7.54 (d, J=8.78 Hz, 1H), 6.77(s, 1H).

Intermediate 41: 3-(methylamino)isoquinoline-7-carboxylic acid, shownbelow, was prepared as follows

Step 1: 7-bromo-N-methylisoquinolin-3-amine

A mixture of 7-bromo-3-chloroisoquinoline (100 mg, 0.4 mmol),methylamine hydrochloride (139 mg, 2.06 mmol), and potassium carbonate(456 mg, 3.30 mmol) in 1-methoxy-2-(2-methoxyethoxyl)ethane (1 mL) washeated to 150° C. and stirred for 60 hours. Additional methylaminehydrochloride (100 mg, 1.5 mmol) and potassium carbonate (200 mg, 1.4mmol) were added and heating was continued for another 40 hours. Thereaction was cooled to room temperature and diluted with water. Themixture was stirred for 30 minutes. The resulting solid was filteredoff, rinsed with water and dried under vacuum. Purification by flashcolumn chromatography (10-30% ethyl acetate/heptanes) gave the titlecompound (82 mg) as a pale yellow solid. −APCI (M−H+1) 237.8; ¹H NMR(400 MHz, CDCl₃, δ): 8.70 (s, 1H), 7.84 (d, J=1.95 Hz, 1H), 7.48 (dd,J=8.97, 2.15 Hz, 1H), 7.38 (d, J=8.97 Hz, 1H), 6.39 (s, 1H), 2.92 (s,3H).

Step 2: 3-(methylamino)isoquinoline-7-carboxylic acid

The title compound was prepared by a method analogous to that describedin Steps 3-4 of Intermediate 21, using7-bromo-N-methylisoquinolin-3-amine. +ESI (M+H) 203.1; ¹H NMR (400 MHz,CD₃OD, δ): 8.87 (s, 1H), 8.51 (s, 1H), 7.98 (dd, J=8.88, 1.66 Hz, 1H),7.58 (d, J=8.78 Hz, 1H), 6.60 (s, 1H), 2.93 (s, 3H).

Intermediate 42: 3-chloro-1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid,shown below, was prepared as follows

A suspension of 1H-pyrrolo[2,3-b]pyridine-5-carboxylic acid (250 mg, 1.5mmol) in N,N-dimethylformamide (5 mL) was warmed to 40° C.N-chlorosuccinimide (243 mg, 1.62 mmol) was added and the mixture wasstirred at 55° C. for 5 hours. The reaction was cooled to roomtemperature and left stirring for 2 days. The mixture was diluted withwater (20 mL) and stirred overnight. The resulting solid was collectedby filtration and dried to give the title compound (161 mg, 55%). +ESI(M+H) 197.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 13.08 (br. s., 1H), 12.39(br. s., 1H), 8.86 (d, J=1.8 Hz, 1H), 8.40 (d, J=1.2 Hz, 1H), 7.84 (d,J=2.5 Hz, 1H).

Intermediate 43: 6-bromo-3-methoxyisoquinoline, shown below, wasprepared as follows

A mixture of 6-bromoisoquinolin-3-ol (606 mg, 2.70 mmol), silvercarbonate (1.5 g, 5.3 mmol), and N,N-dimethylformamide (12 mL) wasstirred at room temperature for 16 minutes. Methyl iodide (186 μL, 2.97mmol) was added and the reaction was left stirring for 18 hours. Thereaction was diluted with methanol and filtered through Celite. Thefiltrate was concentrated and purified by flash column chromatography togive the title compound (90 mg, 14%). +ESI (M+H+1) 240.0; ¹H NMR (400MHz, CDCl₃, δ): 8.91 (s, 1H), 7.86 (d, J=1.8 Hz, 1H), 7.73 (d, J=8.8 Hz,1H), 7.43 (dd, J=8.8, 1.8 Hz, 1H), 6.90 (s, 1H), 4.02 (s, 3H).

Intermediate 44: 2-chloroquinoline-7-carboxylic acid, shown below, wasprepared as follows

Step 1: ethyl 2-chloroquinoline-7-carboxylate

Phosphorus oxychloride (1.94 mL, 20.7 mmol) was added to a solution of7-(ethoxycarbonyl)quinoline 1-oxide (450 mg, 2.07 mmol) indichloromethane (15 mL). The reaction was heated to 50° C. for 3 hours.The reaction was then cooled to room temperature and was slowly pouredinto 200 mL of water, with stirring. The mixture was allowed to stir for1 hour and was then neutralized with 1 N aqueous potassium hydroxide.The mixture was extracted with dichloromethane (3×). The extracts werewashed with brine, dried over magnesium sulfate, filtered, andconcentrated. Purification by column chromatography (0-20% ethylacetate/heptanes) gave the title compound (254 mg, 52%) as a whitesolid. ¹H NMR (400 MHz, CDCl₃, δ): 8.70-8.79 (m, 1H), 8.13-8.18 (m, 2H),7.87 (d, J=8.39 Hz, 1H), 7.47 (d, J=8.58 Hz, 1H), 4.44 (q, J=7.02 Hz,2H), 1.43 (t, J=7.12 Hz, 3H).

Step 2: 2-chloroquinoline-7-carboxylic acid

To a solution of ethyl 2-chloroquinoline-7-carboxylate (800 mg, 3.4mmol) in tetrahydrofuran (10 mL) was added 1 N aqueous lithium hydroxide(7 mL, 7 mmol). The reaction was stirred at room temperature overnight.The reaction was concentrated and the residue was diluted with water andacidified with 1 N aqueous hydrochloric acid. The resulting precipitatewas collected by filtration and dried under vacuum to give the titlecompound (648 mg, 92%) as a white powder. +ESI (M+H) 208.1; ¹H NMR (400MHz, DMSO-d₆, δ): 13.43 (s, 1H), 8.53 (d, J=8.7 Hz, 1H), 8.44-8.45 (m,1H), 8.14 (d, J=8.4 Hz, 1H), 8.07-8.11 (m, 1H), 7.70 (d, J=8.5 Hz, 1H).

Intermediate 44: 2-((2,2,2-trifluoroethyl)amino)quinoline-7-carboxylicacid, shown below, was prepared as follows

Step 1: methyl quinoline-7-carboxylate

The title compound was prepared by a method analogous to that describedin Step 3 of Intermediate 21 using 7-bromoquinoline as the startingmaterial.

Step 2: 7-(methoxycarbonyl)quinoline 1-oxide, shown below, was preparedas follows:

To a solution of methyl quinoline-7-carboxylate (17.8 g, 94.87 mmol) indichloromethane (315 mL) was added peracetic acid (39.9 mL, 190 mmol,32% in acetic acid). The reaction was stirred at room temperatureovernight. Peracetic acid (10 mL, 48 mmol, 32% in acetic acid) was addedand the mixture was stirred for 5 h. The reaction mixture was dilutedwith a saturated solution of aqueous sodium bicarbonate. The aqueousphase was extracted into dichloromethane (2×1 L). The extracts werecombined, dried over magnesium sulfate, filtered and concentrated underreduced pressure. Purification by flash chromatography (2-15% methanolin dichloromethane) gave the title compound (17.4 g, 90%) as a yellowsolid. ¹H NMR (400 MHz, CHLOROFORM-d, δ): 9.41 (1H, s), 8.56 (1H, dd,J=6.0, 0.8 Hz), 8.24 (1H, dd, J=8.5, 1.7 Hz), 7.93 (1H, d, J=8.6 Hz),7.75 (1H, d, J=8.6 Hz), 7.39 (1H, dd, J=8.6, 6.0 Hz), 4.01 (3H, s)

Step 3: methyl 2-((2,2,2-trifluoroethyl)amino)quinoline-7-carboxylate,shown below, was prepared as follows

To a solution of 7-(methoxycarbonyl)quinoline 1-oxide (200 mg, 0.984mmol) and 2,2,2-trifluoroethylamine (292 mg, 0.295 mmol) at 0° C. wasadded 4-methylbenzenesulphonic anhydride (964 mg, 2.95 mmol) portionwiseover a period of 45 minutes. The reaction was allowed to warm up to roomtemperature and stirred overnight. The reaction was diluted withdichloromethane and washed with a saturated solution of ammoniumchloride. The aqueous layer was extracted into dichloromethane (1×). Theorganics were combined and washed with brine, dried over magnesiumsulfate, filtered and concentrated under reduced pressure. Purificationby flash chromatography gave the title compound (172 mg, 62%). +ESI(M+H) 285.1

Step 4: 2-((2,2,2-trifluoroethyl)amino)quinoline-7-carboxylic acid

To a solution of methyl2-((2,2,2-trifluoroethyl)amino)quinoline-7-carboxylate (172 mg, 0.605mmol) in tetrahydrofuran (5 mL) was added aqueous lithium hydroxide(1.82 mL, 1.82 mmol, 1M solution) at room temperature. The reaction wasstirred for 2.5 days. The solvent was removed under reduced pressure andthe residue was acidified with 1N aqueous hydrochloric acid. Theresulting precipitate was filtered and dried to give the title compound(65 mg, 40%) +ESI (M+H) 271.1, ¹H NMR (400 MHz, DMSO-d6, δ): 4.31-4.41(m, 2H) 7.01 (d, J=8.87 Hz, 1H) 7.69-7.80 (m, 2H) 8.06 (d, J=8.66 Hz,1H) 8. 14 (s, 1H) 13.03 (bs, 1H)

Intermediate 45: 2-((2,2-difluoropropyl)amino)quinoline-7-carboxylicacid, shown below, was prepared as follows

The title compound was prepared by a method analogous to that describedfor Intermediate 44, using 2,2-difluoroethylamine instead of2,2,2-trifluoroethylamine. +ESI (M+H) 267.2; ¹H NMR (400 MHz, DMSO-d6,δ): 1.63 (t, J=19.02 Hz, 3H) 3.89-3.99 (m, 2H) 6.97 (d, J=8.97 Hz, 1H)7.54 (t, 1H) 7.62-7.68 (m, 1H) 7.71 (d, J=8.19 Hz, 1H) 7.96 (d, J=9.10Hz, 1H) 8.06-8.09 (m, 1H) 12.95 (bs, 1H).

Intermediate 46: 7-chloro-1H-benzo[d][1,2,3]triazole-5-carboxylic acid,shown below, was prepared as follows

To a solution of 3,4-diamino-5-chlorobenzoic acid (125 mg, 0.67 mmol) inconcentrated sulfuric acid (0.45 mL) was added water (2 mL) at 0° C. Thereaction mixture was stirred at 0° C. for 1 h. The mixture was leftstirring overnight. The reaction was diluted with water and theresulting precipitate was filtered to give the title compound (124 mg,94%) as a brown solid. +APCI (M+H) 198.0; ¹H NMR (400 MHz, METHANOL-d₄,δ): 8.53 (d, J=1.2 Hz, 1H), 8.10 (d, J=1.0 Hz, 1H)

Example 11′-isopropyl-1-(2-methyl-1H-benzo[d]imidazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one

To a solution of 2-methyl-1H-benzo[d]imidazole-5-carboxylic acid (42 mg,0.13 mmol) in dichloromethane (2 mL) was added1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1H)-onehydrochloride salt (42 mg, 0.13 mmol), triethylamine (0.01 mL, 0.07mmol), and (1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate (54.8 mg, 0.144 mmol). The reaction mixture wasstirred at room temperature overnight. The mixture was concentrated invacuo, and the resultant solids were dissolved in ethyl acetate, washedwith saturated sodium bicarbonate and dried over sodium sulfate,filtered, and concentrated in vacuo. The residue was dissolved indimethyl sulfoxide (1 mL) and purified by reversed-phase HPLC (column:Waters XBridge C18 19×100, 5 μm; mobile phase A: 0.03% NH₄OH in water(v/v); mobile phase B: 0.03% NH₄OH in acetonitrile (v/v); gradient: 90%A/10% B linear to 0% A/100% B in 8.5 min, hold at 0% A/100% B for 10.0min; flow: 25 mL/min. +ESI (M+H) 407.2; HPLC retention time 1.74 minutes(Method A)

Example 21-(3,7-dimethyl-1H-indazole-5-carbonyl)-1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one

To a solution of1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7(1′H)-one hydrochloride salt (Intermediate 2, 430 mg, 1.3 mmol) and3,7-dimethyl-1H-indazole-5-carboxylic acid (306 mg, 1.6 mmol) indimethylformamide (2 mL) was added triethylamine (0.75 mL, 5.4 mmol),4-dimethylaminopyridine (33 mg, 0.37 mmol), and 1-propanephosphonic acidcyclic anhydride (0.52 mL, 1.74 mmol, 50% solution in ethyl acetate),and the reaction mixture was stirred overnight at room temperature. Thereaction mixture was concentrated in vacuo, taken up in ethyl acetateand washed with saturated aqueous sodium bicarbonate. The organic layerwas dried over sodium sulfate, filtered, and concentrated to a solid.The solid was purified via flash column chromatography (0-15%methanol/dichloromethane) to afford a glassy solid. The glassy solid wasstirred in ethyl acetate for 16 hours and the resulting solid collectedby vacuum filtration to afford the desired product as a white solid (138mg). +ESI (M+H) 421.0; ¹H NMR (400 MHz, CD₃OD, δ): 7.65 (s, 1H) 7.42 (s,1H) 7.21 (s, 1H) 5.50 (m, 1H) 3.95 (br. s., 1H) 3.50-3.62 (br. s., 3H)2.97 (s, 2H) 2.56 (m, 6H) 1.83 (br. s., 4H) 1.44 (d, 6H).

Example 31′-isopropyl-1-(2-methyl-2H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one

To a solution of 2-methyl-2H-indazole-5-carboxyllic acid (28 mg, 0.16mmol) in dry dimethylformamide was added1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (37 mg, 0.19 mmol) and1-hydroxybenzotriazole (26 mg, 0.19 mmol) N,N-diisopropylethylamine (84μL, 0.48 mmol). The reaction mixture was stirred at room temperature for10 minutes and then1′-isopropyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-onehydrochloride was added (Intermediate 2, 30 mg, 0.12 mmol) and thereaction was stirred for 16 hours. The mixture was poured into chilledwater and the resulting precipitate was collected by vacuum filtration.The obtained solid was triturated from diethyl ether to afford1′-isopropyl-1-(2-methyl-2H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(1′H)-one(25 mg). +ESI (M+H) 407.3; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.41 (s, 1H),7.85 (s, 1H), 7.75 (s, 1H), 7.63 (d, 1H), 7.40 (s, 1H), 7.20 (s, 1H),5.40 (m, 1H), 4.18 (s, 3H), 3.60 (br. s., 4H), 2.85 (s, 2H), 1.70 (br.s., 4H), 1.35 (d, 6H).

The compounds listed in Table 1 below were prepared using proceduresanalogous to those described above for the synthesis of the compounds ofExamples 1-3 using the appropriate starting materials which areavailable commercially, prepared using preparations well-known to thoseskilled in the art, or prepared in a manner analogous to routesdescribed above for other intermediates. The compounds listed below wereisolated initially as the free base and may be converted to apharmaceutically acceptable salt for testing.

TABLE 1

Ex. R₁ R₂ Analytical Data 4 iPr^(a)

+ESI (M + H) 407.3; HPLC retention time 2.16 minutes (Method A) 5 iPr

+ESI (M + H) 393.3; HPLC retention time 2.07 minutes (Method A) 6tBu^(b)

+ESI (M + H) 407.1; ¹H NMR (500 MHz, DMSO-d₆, δ): 13.23 (s, 1 H), 8.14(s, 1 H), 7.83 (d, 2 H), 7.59 (s, 1 H), 7.36 (d, J = 8.5 Hz, 1 H), 7.33(s, 1 H), 3.50 (br. s., 4 H), 2.85 (s, 2 H), 1.64 (m, 13 H). 7 tBu

+ESI (M + H) 421.2; 1H NMR (500 MHz, DMSO-d₆, δ): 13.31 (br. s., 1 H),8.12 (s, 1 H), 7.83 (s, 1 H), 7.62 (s, 1 H), 7.33 (s, 1 H), 7.14 (s, 1H), 3.64 (m, 4 H), 2.85 (s, 2 H), 2.53 (s, 3 H), 1.64 (m, 13 H). 8 tBu

+ESI (M + H) 435.3; ¹H NMR (400 MHz, CDCl₃, δ): 10.34 (m, 1 H), 7.57 (s,1 H), 7.23 (s, 1 H), 7.15 (s, 1 H), 6.59 (m, 1 H), 3.59 (m, 4 H), 2.83(s, 2 H), 2.55 (br. s., 3 H), 2.49 (s, 3 H), 1.86 (br. s., 4 H), 1.70(s, 9 H). 9 iPr

+ESI (M + H) 393.1; ¹H NMR (300 MHz, DMSO-d₆, δ): 11.85 (s, 1 H), 8.25(s, 1 H), 8.00 (s, 1 H), 7.85 (s, 1 H), 7.55 (m, 1 H), 7.40 (s, 1 H),6.50 (m, 1 H), 5.40 (m, 1 H), 3.60 (m, 4 H), 2.85 (s, 2 H), 1.75 (br. s,4 H), 1.35 (d, 6 H). 10 iPr

+ESI (M + H) 393.2; ¹H NMR (400 MHz, DMSO-d₆, δ): 11.55 (s, 1 H), 8.35(s, 1 H), 7.90 (s, 1 H), 7.85 (m, 2 H), 7.40 (s, 1 H), 6.60 (s, 1 H),5.40 (m, 1 H), 3.60 (m, 4 H), 2.85 (s, 2 H), 1.75 (m, 4 H), 1.35 (d, 6H). 11 iPr

+ESI (M + H) 427.2; ¹H NMR (400 MHz, DMSO-d₆, δ): 13.78 (s, 1 H), 8.26(s, 1 H), 7.90 (s, 1 H), 7.80 (s, 1 H), 7.45 (s, 1 H), 7.41 (s, 1 H),5.40 (m, 1 H), 3.60 (m, 4 H), 2.85 (s, 2 H), 1.75 (br. s., 4 H), 1.38(d, 6 H). 12 iPr

+ESI (M + H) 423.2; HPLC retention time 2.19 minutes (Method A) 13 iPr

+ESI (M + H) 407.3; ¹H NMR (300 MHz, DMSO-d₆, δ): 8.15 (s, 1 H), 7.88(s, 1 H), 7.80 (s, 1 H), 7.70 (d, 1 H), 7.40 (m, 2 H), 5.45 (m, 1 H),4.06 (s, 3 H), 2.85 (s, 2 H), 1.70 (br. s., 4 H), 1.35 (d, 6 H). 14 iPr

+ESI (M + H) 421.2; ¹H NMR (300 MHz, DMSO-d₆, δ): 13.35 (s, 1 H), 8.15(s, 1 H), 7.86 (s, 1 H), 7.62 (s, 1 H), 7.41 (s, 1 H), 7.15 (s, 1 H),5.40 (m, 1 H), 3.60 (m, 4 H), 2.95 (m, 4 H), 1.70 (br. s., 4 H), 1.30(m, 9 H). 15 iPr

+ESI (M + H) 421.4; ¹H NMR (400 MHz DMSO-d₆, δ): 12.78 (s, 1 H), 7.85(s, 1 H), 7.78 (s, 1 H), 7.50 (d, 1 H), 7.40 (s, 1 H), 7.36 (d, 1 H),5.40 (m, 1 H), 3.60 (m, 4 H), 2.94 (q, 2 H), 2.85 (s, 2 H), 2.70 (br.s., 4 H), 1.35 (m, 9 H). 16 iPr

+ESI (M + 30H) 407.3; HPLC retention time 2.15 minutes (Method A) 17 iPr

+ESI (M + H) 464.3; ¹H NMR (300 MHz DMSO-d₆, δ): 7.92 (s, 1 H), 7.68 (m,1 H), 7.50 (t, 1 H), 7.41 (s, 1 H), 7.24 (s, 1 H), 5.42 (m, 1 H), 4.10(s, 3 H), 3.92 (s, 1 H), 3.90-3.38 (m, 4 H), 2.86 (s, 2 H), 1.85-1.52(m, 4 H), 1.38 (m, 6 H). 18 iPr

+ESI (M + H) 393.2; ¹H NMR (300 MHz, DMSO-d₆, δ): 11.81 (s, 1 H), 8.38(m, 1 H), 7.95 (s, 1 H), 7.76 (d, 1 H), 7.41 (s, 1 H), 7.19 (m, 1 H),6.85 (s, 1 H), 5.42 (m, 1 H), 4.02-3.58 (m, 4 H), 2.90 (s, 1 H),1.86-1.46 (m, 4 H), 1.39 (d, 6 H). 19 iPr

+ESI (M + H) 394.0; ¹H NMR (300 MHz, DMSO-d₆, δ): 13.56 (s, 1 H), 8.56(s, 1 H), 8.38 (s, 1 H), 8.08 (s, 1 H), 7.94 (s, 1 H), 7.42 (s, 1 H),5.42 (m, 1 H), 3.94-3.41 (m, 4 H), 2.86 (s, 1 H), 1.87-1.60 (m, 4 H),1.35 (m, 6 H). 20 iPr

+ESI (M + H) 404.3; HPLC retention time 2.10 minutes (Method A) 21 iPr

+ESI (M + H) 404.3; HPLC retention time 1.79 minutes (Method A) 22 iPr

+ESI (M + H) 404.3; HPLC retention time 1.75 minutes (Method A) 23 iPr

+ESI (M + H) 404.3; HPLC retention time 1.76 minutes (Method A) 24 iPr

+ESI (M + H) 404.3; HPLC retention time 1.84 minutes (Method A) 25 iPr

+ESI (M + H) 418.3; ¹H NMR (400 MHz, DMSO-d₆, δ): 12.81 (s, 1 H), 7.85(s, 2 H), 7.80 (dd, J = 8.7, 0.9 Hz, 1 H), 7.51 (dd, J = 8.7, 1.5 Hz, 1H), 7.38 (s, 1 H), 5.39 (s, 1 H), 3.32 - 3.87 (m, 4 H), 2.83 (s, 2 H),1.69 (m, 4 H), 1.33 (d, J = 6.4 Hz, 6 H). 26 iPr

+ESI (M + H) 434.4; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.68 (d, J = 2.1 Hz, 1H), 8.26 (d, J = 1.4 Hz, 1 H), 7.87-7.96 (m, 2 H), 7.40-7.47 (m, 2 H),7.38 (s, 1 H), 5.33-5.46 (m, 1 H), 3.88 (s, 3 H), 3.74-3.86 (m, 1 H),3.60-3.74 (m, 1 H), 3.44-3.58 (m, 1 H), 3.31-3.44 (m, 1 H), 2.83 (s, 2H), 1.59-1.82 (m, 4 H), 1.28-1.39 (m, 6 H). 27 iPr

+ESI (M + H) 436.3; ¹H NMR (400 MHz, DMSO-d₆, δ): 13.66 (br. s., 1 H),8.16 (s, 1 H), 7.86 (s, 1 H), 7.75 (br. s., 1 H), 7.62 (d, J = 8.6 Hz, 1H), 7.34-7.39 (m, 3 H), 5.33-5.43 (m, 1 H), 3.36-3.93 (m, 4 H), 2.83 (s,2 H), 1.51-1.78 (m, 4 H), 1.33 (d, J = 6.7 Hz, 6 H). 28 iPr

+ESI (M + H) 434.0; HPLC retention time 2.57 minutes (Method A) 29 iPr

+ESI (M + H) 433.0; HPLC retention time 1.75 minutes (Method A) 30 iPr

+ESI (M + H) 434.2; HPLC retention time 2.48 minutes (Method A) 31 iPr

+ESI (M + H) 427.2; HPLC retention time 2.35 minutes (Method A) 32 iPr

+ESI (M + H) 427.2; HPLC retention time 1.89 minutes (Method A) 33 iPr

+ESI (M + H) 422.2; HPLC retention time 1.66 minutes (Method B) 34 iPr

+ESI (M + H) 427.2; HPLC retention time 2.32 minutes (Method A) 35 iPr

+ESI (M + H) 408.2; HPLC retention time 1.61 minutes (Method B) 36 iPr

+ESI (M + H) 433.2; HPLC retention time 1.89 minutes (Method A) 37 iPr

+ESI (M + H) 422.2; HPLC retention time 1.86 minutes (Method A) 38 iPr

+ESI (M + H) 433.1; HPLC retention time 1.92 minutes (Method A) ^(a)=the term “iPr” is used to designate an isopropyl group ^(b)= the term“tBu” is used to designate a t-butyl group

The compounds listed in Table 2 below were prepared using proceduresanalogous to those described above for the synthesis of the compounds ofExamples 1-3 using the appropriate starting materials which areavailable commercially, prepared using preparations well-known to thoseskilled in the art, or prepared in a manner analogous to routesdescribed above for other intermediates. The compounds listed below wereisolated initially as the free base and may be converted to apharmaceutically acceptable salt for testing.

TABLE 2

Ex. R₁ R₂ Analytical Data 39 iPr^(a)

+APCI (M + H) 393.2; ¹H NMR (400 MHz, CD₃OD, δ): 8.12 (s, 1 H), 7.89 (s,1 H), 7.59 (m, 2 H), 7.44 (dd, J = 8.7, 1.3 Hz, 1 H), 4.55 (m, 1 H),3.76 (m, 4 H), 2.94 (s, 2 H), 1.82 (br. s., 4 H), 1.48 (d, J = 6.6 Hz, 6H). 40 iPr

+APCI (M + H) 407.2; ¹H NMR (400 MHz, CD₃OD, δ): 8.05 (br. s., 1 H),7.54 (m, 2 H), 7.26 (dd, J = 8.4, 1.4 Hz, 1 H), 6.67 (d, J = 6.2 Hz, 1H), 4.55 (m, 1 H), 3.77 (m, 4 H), 2.94 (s, 2 H), 2.57 (s, 3 H), 1.81 (m,4 H), 1.48 (d, J = 6.6 Hz, 6 H). 41 tBu^(b)

+APCI (M + H) 407.2; ¹H NMR (400 MHz, CDCl₃, δ): 8.11 (br. s., 1 H),7.84 (s, 1 H), 7.52 (d, 1 H), 7.44 (dd, 1 H), 7.38 (s, 1 H), 6.22 (s, 1H), 3.51-3.62 (m, 2 H), 2.84 (s, 2 H), 1.73-1.93 (m, 4 H), 1.60 (s, 9H). 42 iPr

+APCI (M + H) 407.3; ¹H NMR (400 MHz, CDCl₃, δ): 11.67 (br. s., 1 H),8.06 (s, 1 H), 7.62 (s, 1 H), 7.24 (d, J = 11.3 Hz, 1 H), 7.18 (d, J =11.3 Hz, 2 H), 6.35 (m, 1 H), 4.53 (m, 1 H), 3.60 (t, J = 10.4 Hz, 2 H),2.81 (s, 2 H), 2.54 (s, 3 H), 1.73 (m, 6 H), 1.49 (d, J = 6.6 Hz, 6 H).43 iPr

+APCI (M + H) 421.3; ¹H NMR (400 MHz, CDCl₃, δ): 10.81 (br. s., 1 H),7.59 (s, 1 H), 7.27 (s, 1 H), 7.18 (s, 1 H), 6.77 (s, 1 H), 4.55 (spt, J= 6.7 Hz, 1 H), 3.62 (t, J = 10.1 Hz, 2 H), 2.84 (s, 2 H), 2.58 (s, 3H), 2.53 (s, 3 H), 1.71-1.94 (m, 4 H), 1.50 (d, 6 H). 44 tBu

+APCI (M + H) 421.0; ¹H NMR (400 MHz, CDCl₃, δ): 11.17 (br. s., 1 H),8.11 (s, 1 H), 7.67 (s, 1 H), 7.40 (s, 1 H), 7.24 (s, 1 H), 6.66 (s, 1H), 3.57 (m, 2 H), 3.67- 4.20 (m, 2 H), 2.86 (s, 2 H), 2.59 (s, 3 H),1.83 (m, 4 H), 1.63 (s, 9 H). 45 tBu

+APCI (M + H) 435.1; ¹H NMR (400 MHz, CDCl₃, δ): 10.93 (br. s., 1 H),7.62 (s, 1 H), 7.40 (s, 1 H), 7.21 (s, 1 H), 6.74 (br. s., 1 H),3.76-4.12 (m, 2 H), 3.64 (t, J = 10.2 Hz, 2 H), 2.86 (s, 2 H), 2.60 (s,3 H), 2.55 (s, 3 H), 1.69-1.87 (m, 4 H), 1.63 (s, 9 H). 46 tBu

+ESI (M + H) 447.2; HPLC retention time 2.73 minutes (Method A) 47t-amyl

+ESI (M + H) 449.2; HPLC retention time 2.35 minutes (Method A) 48 tBu

+APCI (M + H) 448.6; ¹H NMR (400 MHz, CDCl₃, δ): 8.74 (d, J = 2.1 Hz, 1H), 8.10 (d, J = 1.6 Hz, 1 H), 7.96 (d, J = 9.4 Hz, 1 H), 7.37 (dd, J =9.3, 2.8 Hz, 1 H), 7.24 (d, J = 6.4 Hz, 1 H), 7.05 (d, J = 2.7 Hz, 1 H),3.93-4.16 (m, 1 H), 3.89 (s, 3 H), 3.45-3.78 (m, 3 H), 2.82 (s, 2 H),1.60-2.04 (m, 4 H), 1.57 (s, 9 H). 49 tBu

+ESI (M+30H) 447.0; HPLC retention time 2.72 minutes (Method A) 50 tBu

+ESI (M + H) 447.0; HPLC retention time 2.76 minutes (Method A) 51 tBu

+ESI (M + H) 478.0; HPLC retention time 2.24 minutes (Method A) 52 tBu

+ESI (M + H) 407.0; HPLC retention time 2.09 minutes (Method A) 53 tBu

+ESI (M + H) 432.2; HPLC retention time 2.35 minutes (Method A) 54 tBu

+ESI (M + H) 450.2; HPLC retention time 1.95 minutes (Method A) 55 tBu

+ESI (M + H) 448.4; ¹H NMR (400 MHz, CDCl₃, δ): 8.27-8.30 (m, 1 H), 8.04(d, J = 5.9 Hz, 1 H), 7.74- 7.79 (m, 1 H), 7.66-7.70 (m, 1 H), 7.37 (s,1 H), 7.20- 7.24 (m, 1 H), 6.07 (s, 1 H), 4.13 (s, 3 H), 3.56 (br. s., 4H), 2.85 (s, 2 H), 1.62-2.01 (m, 4 H), 1.60 (s, 9 H). 56 tBu

+ESI (M + H) 450.2; HPLC retention time 2.79 minutes (Method A) 57cyclobutyl

+ESI (M + H) 433.1; HPLC retention time 2.14 minutes (Method A) 58 tBu

+ESI (M + H) 433.2; HPLC retention time 1.79 minutes (Method A) 59 tBu

+ESI (M + H) 433.2; HPLC retention time 1.83 minutes (Method B) 60 tBu

+ESI (M + H) 432.2; HPLC retention time 2.27 minutes (Method A) 61 tBu

+ESI (M + H) 422.2; HPLC retention time 1.67 minutes (Method B) 62 tBu

+ESI (M + H) 455.2; HPLC retention time 2.39 minutes (Method A) 63 tBu

+ESI (M + H) 418.2; HPLC retention time 1.9 minutes (Method B) 64 tBu

+ESI (M + H) 441.1; HPLC retention time 2.31 minutes (Method A) 65 tBu

+ESI (M + H) 422.3; ¹H NMR (400 MHz, CD₃OD, δ): 7.78 (d, J = 8.19 Hz, 1H), 7.65 (s, 1 H), 7.34 (s, 1 H), 7.01 (dd, J = 8.29, 1.27 Hz, 1 H),3.87-4.00 (m, 1 H), 3.70-3.82 (m, 1 H), 3.51-3.65 (m, 1 H), 3.40-3.49(m, 1 H), 2.93 (s, 2 H), 1.82-1.93 (m, 2 H), 1.70- 1.79 (m, 2 H), 1.58(s, 9 H). 66 tBu

+ESI (M + H) 437.2; HPLC retention time 2.21 minutes (Method A) 67 tBu

+ESI (M + H) 437.2; HPLC retention time 1.86 minutes (Method A) 68 tBu

+ESI (M + H) 448.2; HPLC retention time 2.08 minutes (Method A) 69 tBu

+ESI (M + H) 447.2; HPLC retention time 1.86 minutes (Method A) 70 tBu

+ESI (M + H) 447.2; HPLC retention time 2.79 minutes (Method A) 71 tBu

+ESI (M + H) 448.2; HPLC retention time 2.55 minutes (Method A) 72 tBu

+ESI (M + H) 433.1; ¹H NMR (400 MHz, DMSO-d₆, δ): 7.88 (m, 1 H), 7.72(br. s., 1 H), 7.62-7.68 (m, 2 H), 7.34 (s, 1 H), 7.07 (m, 1 H), 6.76(d, J = 8.97 Hz, 1 H), 6.53 (br. s., 2 H), 3.30-3.84 (m, 4 H), 2.74-2.83(m, 2 H), 1.55-1.72 (m, 4 H), 1.49 (s, 9 H). 73 tBu

+ESI (M+30H) 447.3; HPLC retention time 1.86 minutes (Method A) 74 tBu

+ESI (M + H) 448.3; HPLC retention time 2.15 minutes (Method A) 75 tBu

+ESI (M + H) 436.2; HPLC retention time 1.81 minutes (Method A) 76 tBu

+ESI (M + H) 447.3; ¹H NMR (400 MHz, DMSO-d₆, δ): 7.82 (d, J = 9.0 Hz, 1H), 7.72 (s, 1 H), 7.60-7.67 (m, 2 H), 7.39-7.43 (m, 1 H), 7.03-7.14 (m,2 H), 6.75 (d, J = 9.0 Hz, 1 H), 3.31-3.86 (m, 4 H), 2.87 (d, J = 4.7Hz, 3 H), 2.79 (5, 2 H), 1.55-1.74 (m, 4 H), 1.49 (s, 9 H). 77 tBu

+ESI (M + H) 422.2; HPLC retention time 1.78 minutes (Method A) 78 tBu

+ESI (M + H) 475.2; HPLC retention time 2.34 minutes (Method A) 79 tBu

+ESI (M + H) 437.0; HPLC retention time 2.27 minutes (Method A) 80 tBu

+ESI (M + H) 475.2; HPLC retention time 2.49 minutes (Method A) 81 tBu

+ESI (M + H) 437.2; HPLC retention time 2.20 minutes (Method A) 82 tBu

+ESI (M + H) 447.4; ¹H NMR (400 MHz, DMSO-d₆, δ): 8.23 (s, 1 H), 7.92(d, J = 5.87 Hz, 1 H), 7.69-7.78 (m, 3 H), 7.57 (dd, J = 8.31, 1.47 Hz,2 H), 6.89 (d, J = 5.87 Hz, 1 H), 3.33-3.85 (m, 4 H), 2.95 (d, J = 4.30Hz, 3 H), 2.82 (s, 2 H), 1.55-1.79 (m, 4 H), 1.52 (s, 9 H). 83 tBu

+ESI (M + H) 475.2; HPLC retention time 2.62 minutes (Method A) 84 tBu

+ESI (M + H) 437.2; HPLC retention time 2.27 minutes (Method A) 85 tBu

+ESI (M + H) 440.1; HPLC retention time 2.53 minutes (Method A) 86 tBu

+ESI (M + H) 440.1; HPLC retention time 2.66 minutes (Method A) 87 tBu

+ESI (M + H) 451.1; HPLC retention time 2.2 minutes (Method A) 88 tBu

+ESI (M + H) 441.1; HPLC retention time 1.9 minutes (Method A) 89 tBu

+ESI (M + H) 436.2; HPLC retention time 1.66 minutes (Method B) 90 tBu

+ESI (M + H) 448.3; HPLC retention time 2.35 minutes (Method A) 91 tBu

+ESI (M + H) 447.2; HPLC retention time 1.86 minutes (Method A) 92 tBu

+ESI (M + H) 448.0; HPLC retention time 2.46 minutes (Method A) 93 tBu

+ESI (M + H) 436.1; HPLC retention time 1.90 minutes (Method A) 94 tBu

+ESI (M + H) 433.1; HPLC retention time 1.73 minutes (Method A) 95 tBu

+ESI (M + H) 447.2; HPLC retention time 1.93 minutes (Method A) 96 tBu

+ESI (M + H) 441.1; HPLC retention time 2.21 minutes (Method A) 97 tBu

+ESI (M + H) 402.8; HPLC retention time 1.91 minutes (Method A) 98 tBu

^(a)= the term “iPr” is used to designate an isopropyl group ^(b)= theterm “tBu” is used to designate a t-butyl group

Example 992′-tert-butyl-1-(7-methoxy-1H-indazole-5-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

To a mixture of2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt (Intermediate 4, 25 mg, 0.075 mmol) and7-methoxy-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylic acid(Intermediate 18, 25 mg, 0.090 mmol) in N,N-dimethylformamide (0.4 mL)was added triethylamine (0.05 mL, 0.37 mmol). The mixture was stirredfor 5 minutes. Then 1-propanephosphonic acid cyclic anhydride (0.09 mL,0.1 mmol, 50% solution in ethyl acetate) was added and the reaction wasstirred at room temperature overnight. The reaction was diluted withwater and extracted with ethyl acetate (3×). The combined organic layerswere washed with brine, dried over sodium sulfate, filtered, andconcentrated to a yellow gum. To this crude material was addedhydrochloric acid (0.19 mL, 0.75 mmol, 4 M in dioxane). The mixture wasstirred at room temperature overnight. The reaction was concentrated.Purification by reversed-phase HPLC gave the title compound (3.4 mg,10%). +ESI (M+H) 437.3; HPLC retention time 2.12 minutes (Method A).

Example 1001-(1-aminoisoquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

Step 1:2′-tert-butyl-1-(1-(4-methoxybenzylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

The title compound was prepared by a method analogous to that describedin Example 3, using2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt (Intermediate 4) and1-(4-methoxybenzylamino)isoquinoline-7-carboxylic acid (Intermediate27). +ESI (M+H) 553.5.

Step 2:1-(1-aminoisoquinoline-7-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

To a solution of2′-tert-butyl-1-(1-(4-methoxybenzylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one(28 mg, 0.051 mmol) in trifluoroacetic acid (0.51 mL) was added anisol(8.3 μL, 0.076 mmol). The reaction was heated to 65° C. and stirred for19 hours. The reaction was concentrated. Purification by reversed-phaseHPLC gave the title compound (7.1 mg, 32%). +ESI (M+H) 433.2; HPLCretention time 1.79 minutes (Method A).

Example 1011-(1-aminoisoquinoline-6-carbonyl)-2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

The title compound was prepared by a method analogous to that describedfor Example 100, using 1-(4-methoxybenzylamino)isoquinoline-6-carboxylicacid (Intermediate 30) in Step 1. +ESI (M+H) 433.2; HPLC retention time1.82 minutes (Method A).

Example 1022′-tert-butyl-1-(3-methoxyisoquinoline-6-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

To a solution of 6-bromo-3-methoxyisoquinoline (Intermediate 43, 89.9mg, 0.378 mmol) in 1,4-dioxane (6 mL) was added2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt (Intermediate 4, 244 mg, 0.727 mmol) and sodiumacetate (130 mg, 1.5 mmol). Nitrogen gas was bubbled through the mixturefor 15 minutes. Then added[1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium(II), complexwith dichloromethane (102 mg, 0.125 mmol), capped the reaction vesseland bubbled through carbon monoxide gas for 5 minutes. The reaction wasthen heated to 80° C. for 18 hours. The reaction was cooled to roomtemperature and diluted with ethyl acetate. The mixture was filteredthrough Celite and the filtrate was concentrated. Purification byreversed-phase HPLC gave the title compound. +ESI (M+H) 448.1; HPLCretention time 2.26 minutes (Method A).

Example 1032′-tert-butyl-1-(1-(dimethylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-oneStep 1:2′-tert-butyl-1-(1-chloroisoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

The title compound was prepared by a method analogous to that describedfor Example 2, using2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt (Intermediate 4) and1-chloroisoquinoline-7-carboxylic acid, and omitting4-dimethylaminopyridine. +ESI (M+H) 452.3; ¹H NMR (400 MHz, CDCl₃, δ):8.37 (s, 1H), 8.32 (d, J=5.7 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.75-7.79(m, 1H), 7.62 (d, J=5.7 Hz, 1H), 7.39 (s, 1H), 6.42 (s, 1H), 3.43-3.73(m, 4H), 2.87 (s, 2H), 1.64-2.01 (m, 4H), 1.61 (s, 9H).

Step 2:2′-tert-butyl-1-(1-(dimethylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

A solution of dimethylamine in methanol (1.75 mL, 3.50 mmol, 2 M) wasadded to2′-tert-butyl-1-(1-chloroisoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one(158 mg, 0.350 mmol). The reaction vessel was sealed and the mixture washeated to 60° C. and stirred for 65 hours. The reaction was cooled toroom temperature and concentrated. Purification by flash columnchromatography (1-15% methanol/dichloromethane) gave the title compound(99 mg, 61%) as a white solid. +APCI (M+H) 461.4; ¹H NMR (400 MHz,CDCl₃, δ): 8.16-8.20 (m, 1H), 8.12 (d, J=5.9 Hz, 1H), 7.75 (d, J=8.2 Hz,1H), 7.58-7.64 (m, 1H), 7.37 (s, 1H), 7.14 (d, J=5.9 Hz, 1H), 6.00 (br.s., 1H), 3.40-3.71 (m, 4H), 3.10-3.28 (m, 6H), 2.85 (s, 2H), 1.64-1.99(m, 4H), 1.60 (s, 9H).

Example 1042′-tert-butyl-1-(2-chloroquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

The title compound was prepared by a method analogous to that describedfor Example 3 using2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-onehydrochloride salt (Intermediate 4) and 2-chloroquinoline-7-carboxylicacid (Intermediate 44). +ESI (M+H) 452.3; ¹H NMR (400 MHz, CDCl₃, δ):8.12 (d, J=8.2 Hz, 1H), 7.98 (br. s., 1H), 7.88 (dd, J=8.4 Hz, 1H), 7.61(dd, J=8.4, 1.6 Hz, 1H), 7.44 (d, J=8.6 Hz, 1H), 7.39 (s, 1H), 5.91 (br.s., 1H), 4.06-4.22 (m, 1H), 3.38-3.64 (m, 3H), 2.85 (br. s., 2H),1.67-1.97 (m, 4H), 1.61 (s, 9H).

Example 1052′-tert-butyl-1-(2-(dimethylamino)quinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

A solution of dimethylamine in tetrahydrofuran (2.2 mL, 4.4 mmmol, 2.0M) was added to2′-tert-butyl-1-(2-chloroquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one(100 mg, 0.2 mmol). The reaction vessel was sealed and the mixture washeated to 70° C. for 15 hours. The reaction was cooled to roomtemperature and concentrated. Purification by reversed-phase HPLC gavethe title compound (25 mg, 25%). +ESI (M+H) 461.2; HPLC retention time1.96 minutes (Method A).

The compounds listed in Table 3 below were prepared using a procedureanalogous to that described above for the synthesis of the Example 105using the appropriate starting materials which are availablecommercially, prepared using preparations well-known to those skilled inthe art, or prepared in a manner analogous to routes described above forother intermediates. The compounds listed below were isolated initiallyas the free base and may be converted to a pharmaceutically acceptablesalt for testing.

TABLE 3

Ex. R Analytical Data 106

+ESI (M + H) 473.1; HPLC retention time 2.21 minutes (Method B). 107

+ESI (M + H) 475.2; HPLC retention time 1.82 minutes (Method A) 108

+ESI (M + H) 475.4; ¹H NMR (400 MHz, CDCl₃, δ): 7.82 (d, J = 8.76 Hz, 1H), 7.56-7.66 (m, 2 H), 7.37 (s, 1 H), 7.13-7.18 (m, 1 H), 6.89 (d, J =9.15 Hz, 1 H), 5.79 (s, 1 H), 4.02-4.15 (m, 1 H), 3.68 (m, 2 H),3.41-3.62 (m, 3 H), 3.17-3.22 (m, 3 H), 2.84 (s, 2 H), 1.66-1.93 (m, 4H), 1.61 (s, 9 H), 1.21 (m, 3 H). 109

+APCI (M + H) 487.6; ¹H NMR (400 MHz, CDCl₃, δ): 7.80 (d, J = 8.91 Hz, 1H), 7.58-7.61 (m, 2 H), 7.37 (s, 1 H), 7.19 (dd, J = 9.76, 1.53 Hz, 1H), 6.64 (d, J = 8.95 Hz, 1 H), 5.81 (s, 1 H), 4.98-5.04 (m, 1 H),4.39-4.49 (m, 1 H), 4.02-4.16 (m, 1 H), 3.40-3.64 (m, 3 H), 2.83 (s, 2H), 2.43-2.52 (m, 2 H), 1.72-1.96 (m, 8 H), 1.61 (s, 9 H). 110

+APCI (M + H) 475.6; ¹H NMR (400 MHz, CDCl₃, δ): 7.79 (d, J = 8.81 Hz, 1H), 7.61-7.62 (m, 1 H), 7.59 (d, J = 8.21 Hz, 1 H), 7.37 (s, 1 H), 7.19(dd, J = 8.17, 1.57 Hz, 1 H), 6.65 (d, J = 8.81 Hz, 1 H), 5.79 (s, 1 H),4.69-4.73 (m, 1 H), 4.00-4.15 (m, 1 H), 3.41-3.63 (m, 5 H), 2.83 (s, 2H), 1.64-1.92 (m, 6 H), 1.61 (s, 9 H), 1.02 (t, J = 7.41 Hz, 3 H). 111

+APCI (M + H) 491.6; ¹H NMR (400 MHz, CDCl₃, δ): 7.77 (d, J = 8.91 Hz, 1H), 7.63-7.64 (m, 1 H), 7.59 (d, J = 8.13 Hz, 1 H), 7.37 (s, 1 H), 7.19(dd, J = 8.20, 1.56 Hz, 1 H), 6.65 (d, J = 8.78 Hz, 1 H), 5.79 (s, 1 H),5.05-5.10 (m, 1 H), 4.03-4.13 (m, 1 H), 3.70-3.73 (m, 2 H), 3.63 (t, 2H), 3.43-3.61 (m, 3 H), 3.40 (s, 3 H), 2.84 (s, 2 H), 1.66-1.93 (m, 4H), 1.61 (s, 9 H). 112

+APCI (M + H) 461.5; ¹H NMR (400 MHz, CDCl₃, δ): 7.79 (d, J = 8.82 Hz, 1H), 7.62-7.63 (m, 1 H), 7.59 (d, J = 8.21 Hz, 1 H), 7.37 (s, 1 H), 7.19(dd, J = 8.19 Hz, 1.57 Hz, 1 H), 6.64 (d, J = 8.81 Hz, 1 H), 5.78 (s, 1H), 4.67-4.71 (m, 1 H), 4.00-4.15 (m, 1 H), 3.44- 3.59 (m, 5 H), 2.84(s, 2 H), 1.64-1.94 (m, 4 H), 1.61 (s, 9 H), 1.29 (t, J = 7.22 Hz, 3 H).113

+ESI (M + H) 489.3; HPLC retention time 1.8 minutes (Method A)

The compounds listed in Table 4 below were prepared using proceduresanalogous to those described above for the synthesis of the compounds ofExamples 1-3 using the appropriate starting materials which areavailable commercially, prepared using preparations well-known to thoseskilled in the art, or prepared in a manner analogous to routesdescribed above for other intermediates. The compounds listed below wereisolated initially as the free base and may be converted to apharmaceutically acceptable salt for testing.

TABLE 4

Ex. R₁ R₂ Analytical Data 114 tBu^(a)

+ESI (M + H) 461.2; ¹H NMR (400 MHz, CDCl₃, δ): 7.68 (s, 1 H), 7.34 (s,1 H), 7.25 (s, 1 H), 6.23 (s, 1 H), 4.85 (br. s., 1 H), 4.26 (br. s., 1H), 2.64-2.78 (m, 2 H), 2.61 (s, 3 H), 2.56 (s, 3 H), 1.74-2.36 (m, 8H), 1.59 (s, 9 H). ^(a)= the term “tBu” is used to designate a t-butylgroup

The compounds listed in Table 5 below were prepared using a procedureanalogous to that described above for the synthesis of the Example 103using the appropriate starting materials which are availablecommercially, prepared using preparations well-known to those skilled inthe art, or prepared in a manner analogous to routes described above forother intermediates. The compounds listed below were isolated initiallyas the free base and may be converted to a pharmaceutically acceptablesalt for testing.

TABLE 5

Ex. R Analytical Data 115

+ESI (M + H) 473.3; HPLC retention time 2.01 minutes (Method A). 116

+ESI (M + H) 475.3; HPLC retention time 2.06 minutes (Method A) 117

+ESI (M + H) 475.3; HPLC retention time 1.99 minutes (Method A) 118

+ESI (M + H) 487.3; HPLC retention time 2.12 minutes (Method A) 119

+ESI (M + H) 475.3; HPLC retention time 2.08 minutes (Method A) 120

+ESI (M + H) 491.3; HPLC retention time 1.99 minutes (Method A) 121

+ESI (M + H) 461.2; HPLC retention time 1.98 minutes (Method A) 122

+ESI (M + H) 489.3; HPLC retention time 1.85 minutes (Method A)

Example 1232′-(tert-butyl)-1-(1-(tert-butylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

Step 1: 1-chloroisoquinoline-7-carboxylic acid

To a suspension of 7-bromo-1-chloroisoquinoline (2.000 g, 8.247 mmoles)in THF (12 mL) and diethyl ether (12 mL) cooled down to −78° C. wasadded n-BuLi (3.96 mL, 9.9 mmol, 2.5M in hexanes). Stirred for fiveminutes and then bubbled carbon dioxide while venting with a needle forapproximately one minute. The reaction mixture was warmed up to 0° C.and 15 mL of a 1N aqueous sodium hydroxide were added. The mixture wasdiluted with diethyl ether stirred for 18 h. The organic and aqueouslayers were separated and the organics were washed with 1N aqueoussodium hydroxide and water. The aqueous fractions were combined andacidified to pH 4 with 1N aqueous hydrochloric acid. The resultingsolids were collected by filtration and dried to give the title compound(1.252 g, 73%). +ESI (M+H) 208.1 ¹H NMR (400 MHz, DMSO-d₆) d ppm 13.58(br. s., 1H) 8.86 (m, 1H) 8.43 (d, J=5.67 Hz, 1H) 8.33 (dd, J=8.61, 1.57Hz, 1H) 8.19 (d, J=8.41 Hz, 1H) 8.01 (dd, 1H)

Step 2:2′-(tert-butyl)-1-(1-(tert-butylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one

To a suspension of 1-chloroisoquinoline-7-carboxylic acid (100 mg, 0.482mmol), RuPhos (6.5 mg, 0.014 mmol), BrettPhos (11.2 mg, 0.014 mmol) andsodium tert-butoxide (70.2 mg, 0.723 mmol) in dioxane (0.5 mL) was addedt-butylamine (0.254 mL, 2.41 mmol). The vessel was sealed and mixturewas heated to 110° C. and stirred overnight. The reaction was cooleddown to room temperature and lithium bistrimethylsilylamide (0.136 mL,0.723 mmol) was added. The reaction mixture was heated to 110° C. andleft stirring overnight. The reaction mixture was cooled down to roomtemperature and filtered through celite and rinsed with methanol. Thefiltrate was concentrated under reduced pressure and 1N aqueous sodiumhydroxide (1 mL) was added. Partitioned between ethyl acetate and amixture of water and 1N aqueous sodium hydroxide. The layers wereseparated and the aqueous layer was acidified to pH 4. The aqueous layerwas extracted into ethyl acetate. The extracts were dried over magnesiumsulfate, filtered and concentrated under reduced pressure to obtain1-(tert-butylamino)isoquinoline-7-carboxylic acid.

To a suspension of 1-(tert-butylamino)isoquinoline-7-carboxylic acid(24.7 mg, 0.101 mmol) and2′-tert-butyl-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridine]-7′(2′H)-onehydrochloride salt (33.9 mg, 0.101 mmol) in N,N-dimethylformamide (1 mL)was added triethylamine (0.07 mL, 0.50 mmol). The reaction mixture wasstirred at room temperature for 10 minutes. Then 1-propanephosphonicacid cyclic anhydride (0.07 mL, 0.12 mmol, 50% solution in ethylacetate) was added and the reaction mixture was stirred at roomtemperature overnight. N,N-dimethylformamide was removed under reducedpressure and the residue was purified by reversed-phase HPLC to give2′-(tert-butyl)-1-(1-(tert-butylamino)isoquinoline-7-carbonyl)-4′,6′-dihydrospiro[piperidine-4,5′-pyrazolo[3,4-c]pyridin]-7′(2′H)-one(6.1 mg, 24%). +ESI (m+H) 489.3; HPLC retention time 2.94 minutes(Method B).

The compounds listed in Table 6 below were prepared using proceduresanalogous to those described above for the synthesis of the compounds ofExamples 1-3 using the appropriate starting materials which areavailable commercially, prepared using preparations well-known to thoseskilled in the art, or prepared in a manner analogous to routesdescribed above for other intermediates. The compounds listed below wereisolated initially as the free base and may be converted to apharmaceutically acceptable salt for testing.

TABLE 6

Ex. R Analytical Data 124

+ESI (M + H) 515.3; HPLC retention time 2.08 minutes (Method A) 125

+ESI (M + H) 511.3; HPLC retention time 2.10 minutes (Method A)

PHARMACOLOGICAL DATA

Biological Protocols

The utility of the compounds of present invention, in the treatment ofdiseases (such as are detailed herein) in animals, particularly mammals(e.g., humans) may be demonstrated by the activity thereof inconventional assays known to one of ordinary skill in the art, includingthe in vitro and in vivo assays described below. Such assays alsoprovide a means whereby the activities of the compound of the presentinvention can be compared with the activities of other known compounds.

Direct Inhibition of the Activities of ACC1 and ACC2

The ACC inhibitory activity of the compound of the present invention wasdemonstrated by methods based on standard procedures. For example directinhibition of ACC activity, for the compound of Formula (I) wasdetermined using preparations of recombinant human ACC1 (rhACC1) andrecombinant human ACC2 (rhACC2). Representative sequences of therecombinant human ACC1 and ACC2 that can be used in the assay areprovided in FIG. 1 (SEQ ID NO. 1) and FIG. 2 (SEQ. ID NO. 2),respectively.

[1] Preparation of rhACC1. Two liters of SF9 cells, infected withrecombinant baculovirus containing full length human ACC1 cDNA, weresuspended in ice-cold lysis buffer (25 mM Tris, pH 7.5; 150 mM NaCl; 10%glycerol; 5 mM imidazole (EMD Bioscience; Gibbstown, N.J.); 2 mM TCEP(BioVectra; Charlottetown, Canada); Benzonase nuclease (10000 U/100 gcell paste; Novagen; Madison, Wis.); EDTA-free protease inhibitorcocktail (1 tab/50 mL; Roche Diagnostics; Mannheim, Germany). Cells werelysed by 3 cycles of freeze-thaw and centrifuged at 40,000×g for 40minutes (4° C.). Supernatant was directly loaded onto a HisTrap FF crudecolumn (GE Healthcare; Piscataway, N.J.) and eluted with an imidazolegradient up to 0.5 M over 20 column volumes (CV). ACC1-containingfractions were pooled and diluted 1:5 with 25 mM Tris, pH 7.5, 2 mMTCEP, 10% glycerol and direct loaded onto a CaptoQ (GE Healthcare)column and eluted with an NaCl gradient up to 1 M over 20 CV's.Phosphate groups were removed from purified ACC1 by incubation withlambda phosphatase (100 U/10 μM target protein; New England Biolabs;Beverly, Mass.) for 14 hours at 4° C.; okadaic acid was added (1 μMfinal concentration; Roche Diagnostics) to inhibit the phosphatase.Purified ACC1 was exchanged into 25 mM Tris, pH 7.5, 2 mM TCEP, 10%glycerol, 0.5 M NaCl by 6 hour dialysis at 4° C. Aliquots were preparedand frozen at −80° C.

[2] Measurement of rhACC1 inhibition. hACC1 was assayed in a Costar#3676 (Costar, Cambridge, Mass.) 384-well plate using the TranscreenerADP detection FP assay kit (Bellbrook Labs, Madison, Wis.) using themanufacturer's recommended conditions for a 50 μM ATP reaction. Thefinal conditions for the assay were 50 mM HEPES, pH 7.2, 10 mM MgCl₂,7.5 mM tripotassium citrate, 2 mM DTT, 0.1 mg/mL BSA, 30 μM acetyl-CoA,50 μM ATP, and 10 mM KHCO₃ Typically, a 10 μl reaction was run for 120min at 25° C., and 10 μl of Transcreener stop and detect buffer wasadded and the combination incubated at room temp for an additional 1hour. The data was acquired on a Envision Fluorescence reader(PerkinElmer) using a 620 excitation Cy5 FP general dual mirror, 620excitation Cy5 FP filter, 688 emission (S) and a 688 (P) emissionfilter.

[3] Preparation of rhACC2. Human ACC2 inhibition was measured usingpurified recombinant human ACC2 (hrACC2). Briefly, a full length Cytomaxclone of ACC2 was purchased from Cambridge Bioscience Limited and wassequenced and subcloned into PCDNA5 FRT TO-TOPO (Invitrogen, Carlsbad,Calif.). The ACC2 was expressed in CHO cells by tetracycline inductionand harvested in 5 liters of DMEM/F12 with glutamine, biotin, hygromycinand blasticidin with 1 μg/mL tetracycline (Invitrogen, Carlsbad,Calif.). The conditioned medium containing ACC2 was then applied to aSoftlink Soft Release Avidin column (Promega, Madison, Wis.) and elutedwith 5 mM biotin. 4 mgs of ACC2 were eluted at a concentration of 0.05mg/mL (determined by A280) with an estimated purity of 95% (determinedby A280). The purified ACC2 was dialyzed in 50 mM Tris, 200 mM NaCl, 4mM DTT, 2 mM EDTA, and 5% glycerol. The pooled protein was frozen andstored at −80° C., with no loss of activity upon thawing. Formeasurement of ACC2 activity and assessment of ACC2 inhibition, testcompounds were dissolved in DMSO and added to the rhACC2 enzyme as a 5×stock with a final DMSO concentration of 1%.

[4] Measurement of human ACC2 inhibition. hACC2 was assayed in a Costar#3676 (Costar, Cambridge, Mass.) 384-well plate using the TranscreenerADP detection FP assay kit (Bellbrook Labs, Madison, Wis.) using themanufacturer's recommended conditions for a 50 uM ATP reaction. Thefinal conditions for the assay were 50 mM HEPES, pH 7.2, 5 mM MgCl₂, 5mM tripotassium citrate, 2 mM DTT, 0.1 mg/mL BSA, 30 μM acetyl-CoA, 50μM ATP, and 8 mM KHCO₃ Typically, a 10 μl reaction was run for 50 min at25° C., and 10 μl of Transcreener stop and detect buffer was added andthe combination incubated at room temp for an additional 1 hour. Thedata was acquired on an Envision Fluorescence reader (PerkinElmer) usinga 620 excitation Cy5 FP general dual mirror, 620 excitation Cy5 FPfilter, 688 emission (S) and a 688 (P) emission filter.

The results using the recombinant hACC1 and recombinant hACC2Transcreener assays described above are summarized in the table belowfor the Compounds of Formula (I) exemplified in the Examples above.

hACC1 IC50 hACC2 IC50 Example (nM) n (nM) n 1 417 3 312 3 2 24.2 8 19 83 3920 3 2280 3 4 43 6 38 6 5 227 4 174 4 6 269 3 305 3 7 62 3 47 3 8 273 18 3 9 648 3 560 3 10 403 3 498 3 11 88 3 72 3 12 35 3 37 3 13 2040 31550 3 14 131 3 91 3 15 251 3 143 3 16 118 3 90 3 17 72 3 31 3 18 657 3719 3 19 1480 4 2010 4 20 336 3 299 3 21 619 3 341 3 22 1250 3 843 3 23621 3 520 3 24 688 3 320 3 25 259 3 137 3 26 121 3 60 3 27 296 3 132 328 158 4 56 4 29 33 3 9.2 3 30 98 3 30 3 31 179 3 96 3 32 154 3 56 3 33523 3 108 3 34 116 3 50 3 35 487 3 91 3 36 67 3 28 3 37 302 3 127 3 38364 3 107 3 39 329 4 97 4 40 656 4 177 4 41 164 3 67 3 42 68 3 29 3 4329 4 11 4 44 26 3 10 3 45 6.6 7 2.1 7 46 36 3 5.3 3 47 21 7 2.7 7 48 303 9.2 3 49 8.5 3 3.8 3 50 30 3 7.5 3 51 15 3 4.4 3 52 88 3 17 3 53 47 313 3 54 21 3 5.1 3 55 14 4 2.6 4 56 7.4 3 1.9 3 57 50 3 13 3 58 149 3 123 59 123 3 14 3 60 43 3 14 3 61 94 3 13 3 62 8.4 3 2.1 3 63 160 3 28 364 23 4 4.0 4 65 256 3 56 3 66 17 3 2.4 3 67 1340 3 311 3 68 221 3 68 369 185 3 28 3 70 5.8 3 2.4 3 71 20 4 5.2 4 72 28 4 5.9 4 73 48 4 5.5 474 18 3 3.2 3 75 79 3 11 3 76 5.6 5 1.7 5 77 80 3 8.4 3 78 155 3 34 3 79216 3 18 3 80 78 3 21 3 81 46 3 5.3 3 82 21 3 6.4 3 83 163 3 19 3 84 1943 33 3 85 27 3 5.9 3 86 18 3 4.1 3 87 17 3 2.1 3 88 19 4 4.2 4 89 343 344 3 90 209 3 31 3 91 240 3 51 3 92 244 3 55 3 93 65 3 8.6 3 94 130 3 233 95 105 3 9.2 3 96 62 3 7.1 3 97 219 3 45 3 98 99 23 1 5.6 1 100 62 321 3 101 160 3 26 3 102 78 3 12 3 103 73 4 8.9 4 104 89 3 16 3 105 7.4 31.5 3 106 7.5 3 2.6 3 107 6.1 3 2.2 3 108 15 3 2.4 3 109 3.4 3 1.0 3 1105.6 3 1.1 3 111 27 3 3.3 3 112 8.9 3 1.8 3 113 507 3 44 3 114 4820 3 6123 115 29 4 2.9 4 116 20 4 4.7 4 117 332 4 54 4 118 17 4 2.7 4 119 24 42.5 4 120 47 4 6.1 4 121 17 4 2.7 4 122 494 4 77 4 123 78 1 6.3 1 1246.5 3 1.72 3 125 9.0 3 1.7 3 “n” is used to designate the number ofassay runs

Sequence Listing 1 provides a sequence of recombinant human ACC1 (SEQ.ID NO. 1) that can be employed in the Transcreener in vitro assay.

Sequence Listing 2 provides a sequence of recombinant human ACC2 (SEQ.ID NO. 2) that can be employed in the Transcreener in vitro assay.

Acute In Vivo Assessment of ACC Inhibition in Experimental Animals

The ACC inhibitory activity of the compounds of the present inventioncan be confirmed in vivo by evaluation of their ability to reducemalonyl-CoA levels in liver and muscle tissue from treated animals.

Measurement of malonyl-CoA production inhibition in experimental animalscan be determined using the following methodology.

In this method, male Sprague-Dawley Rats, maintained on standard chowand water ad libitum (225-275 g), were randomized prior to the study.Animals were either fed, or fasted for 18 hours prior to the beginningof the experiment. Two hours into the light cycle the animals wereorally dosed with a volume of 5 mL/kg, (0.5% methyl cellulose; vehicle)or with the appropriate compound (prepared in vehicle). Fed vehiclecontrols were included to determine baseline tissue malonyl-CoA levelswhile fasted animals were included to determine the effect fasting hadon malonyl-CoA levels. One hour after compound administration theanimals were asphyxiated with CO₂ and the tissues were removed.Specifically, blood was collected by cardiac puncture and placed into BDMicrotainer tubes containing EDTA (BD Biosciences, N.J.), mixed, andplaced on ice. Plasma was used to determine drug exposure. Liver andquadriceps were removed, immediately freeze-clamped, wrapped in foil andstored in liquid nitrogen.

Tissues were pulverized under liquid N₂ to ensure uniformity insampling. Malonyl-CoA was extracted from the tissue (150-200 mg) with 5volumes 10% tricarboxylic acid in Lysing Matrix A (MP Biomedicals, PN6910) in a FastPrep FP120 (Thermo Scientific, speed=5.5; for 45seconds). The supernatant containing malonyl-CoA was removed from thecell debris after centrifugation at 15000×g for 30 minutes (EppendorfCentrifuge 5402). Samples were stably frozen at −80C until analysis wascompleted.

Analysis of malonyl CoA levels in liver and muscle tissue can beevaluated using the following methodology.

The method utilized the following materials: Malonyl-CoA tetralithiumsalt and malonyl-¹³C₃—CoA trilithium salt which were purchased fromIsotec (Miamisburg, Ohio, USA), sodium perchlorate (Sigma, cat no.410241), trichloroacetic acid (ACROS, cat no. 42145), phosphoric acid(J. T. Baker, cat no. 0260-01), ammonium formate (Fluka, cat no. 17843),methanol (HPLC grade, J. T. Baker, cat no. 9093-33), and water (HPLCgrade, J. T. Baker, 4218-03) were used to make the necessary mobilephases. Strata-X on-line solid phase extraction columns, 25 μm, 20mm×2.0 mm I.D (cat no. 00M-S033-B0-CB) were obtained from Phenomenex(Torrance, Calif., USA). SunFire C18 reversed-phase columns, 3.5 μm, 100mm×3.0 mm I.D. (cat no. 186002543) were purchased from WatersCorporation (Milford, Mass., USA).

This method may be performed utilizing the following equipment.Two-dimensional chromatography using an Agilent 1100 binary pump, anAgilent 1100 quaternary pump and two Valco Cheminert 6-port two positionvalves. Samples were introduced via a LEAP HTC PAL auto sampler withPeltier cooled stack maintained at 10° C. and a 20 μL sampling loop. Theneedle wash solutions for the autosampler were 10% trichloroacetic acidin water (w/v) for Wash 1 and 90:10 methanol:water for Wash 2. Theanalytical column (Sunfire) was maintained at 35° C. using a MicroTechScientific Micro-LC Column Oven. The eluent was analyzed on an ABI SciexAPI3000 triple quadrupole mass spectrometer with Turbo Ion Spray.

Two-dimensional chromatography was performed in parallel using distinctgradient elution conditions for on-line solid phase extraction andreversed-phase chromatography. The general design of the method was suchthat the first dimension was utilized for sample clean-up and capture ofthe analyte of interest followed by a brief coupling of both dimensionsfor elution from the first dimension onto the second dimension. Thedimensions were subsequently uncoupled allowing for gradient elution ofthe analyte from the second dimension for quantification whilesimultaneously preparing the first dimension for the next sample in thesequence. When both dimensions were briefly coupled together, the flowof the mobile phase in the first dimension was reversed for analyteelution on to the second dimension, allowing for optimal peak width,peak shape, and elution time.

The first dimension of the HPLC system utilized the Phenomenex strata-Xon-line solid phase extraction column and the mobile phase consisted of100 mM sodium perchlorate/0.1% (v/v) phosphoric acid for solvent A andmethanol for solvent B.

The second dimension of the HPLC system utilized the Waters SunFire C18reversed-phase column and the mobile phase consisted of 100 mM ammoniumformate for solvent A and methanol for solvent B. The initial conditionof the gradient was maintained for 2 minutes and during this time theanalyte was transferred to the analytical column. It was important thatthe initial condition was at a sufficient strength to elute the analytefrom the on-line SPE column while retaining it on the analytical.Afterwards, the gradient rose linearly to 74.5% A in 4.5 minutes beforea wash and re-equilibration step.

Mass spectrometry when coupled with HPLC can be a highly selective andsensitive method for quantitatively measuring analytes in complexmatrices but is still subject to interferences and suppression. Bycoupling a two dimensional HPLC to the mass spectrometer, theseinterferences were significantly reduced. Additionally, by utilizing theMultiple Reaction Monitoring (MRM) feature of the triple quadrupole massspectrometer, the signal-to-noise ratio was significantly improved.

For this assay, the mass spectrometer was operated in positive ion modewith a TurbolonSpray voltage of 2250V. The nebulizing gas was heated to450° C. The Declustering Potential (DP), Focusing Potential (FP), andCollision Energy (CE) were set to 60, 340, and 42 V, respectively.Quadrupole 1 (Q1) resolution was set to unit resolution with Quadrupole3 (Q3) set to low. The CAD gas was set to 8. The MRM transitionsmonitored were for malonyl CoA: 854.1→347.0 m/z (L. Gao et al. (2007) J.Chromatogr. B 853,303-313); and for malonyl-¹³C₃—CoA: 857.1→350.0 m/zwith dwell times of 200 ms. The eluent was diverted to the massspectrometer near the expected elution time for the analyte, otherwiseit was diverted to waste to help preserve the source and improverobustness of the instrumentation. The resulting chromatograms wereintegrated using Analyst software (Applied Biosystems). Tissueconcentrations for malonyl CoA were calculated from a standard curveprepared in a 10% solution of trichloroacetic acid in water.

Samples comprising the standard curve for the quantification ofmalonyl-CoA in tissue extracts were prepared in 10% (w/v)trichloroacetic acid (TCA) and ranged from 0.01 to 1 μmol/μL.Malonyl-¹³C₃—CoA (final concentration of 0.4 μmol/μL) was added to eachstandard curve component and sample as an internal standard.

Six intra-assay quality controls were prepared; three from a pooledextract prepared from fasted animals and three from a pool made from fedanimals. These were run as independent samples spiked with 0, 0.1 or 0.3μmol/μL ¹²C-malonyl-CoA as well as malonyl-¹³C₃—CoA (0.4 μmol/μL). Eachintra-assay quality control contained 85% of aqueous tissue extract withthe remaining portion contributed by internal standard (0.4 μmol/μL) and¹²C-malonyl-CoA. Inter assay controls were included in each run; theyconsist of one fasted and one fed pooled sample of quadriceps and/or onefasted and one fed pooled sample of liver. All such controls are spikedwith malonyl-¹³C₃—CoA (0.4 μmol/μL).

All publications, including but not limited to issued patents, patentapplications, and journal articles, cited in this application are eachherein incorporated by reference in their entirety.

Although the invention has been described above with reference to thedisclosed embodiments, those skilled in the art will readily appreciatethat the specific experiments detailed are only illustrative of theinvention. It should be understood that various modifications can bemade without departing from the spirit of the invention. Accordingly,the invention is limited only by the following claims.

What is claimed is:
 1. A compound of structure

or a pharmaceutically acceptable salt thereof.
 2. A compound ofstructure


3. A pharmaceutical composition comprising a therapeutically effectiveamount of a compound of claim 1 or a pharmaceutically acceptable saltthereof and a pharmaceutically acceptable excipient, diluent, orcarrier.